US20110159305A1 - Intermediates And Methods For Forming Passivated Surfaces On Oxide Layers And Articles Produced Thereby - Google Patents

Intermediates And Methods For Forming Passivated Surfaces On Oxide Layers And Articles Produced Thereby Download PDF

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US20110159305A1
US20110159305A1 US13/044,759 US201113044759A US2011159305A1 US 20110159305 A1 US20110159305 A1 US 20110159305A1 US 201113044759 A US201113044759 A US 201113044759A US 2011159305 A1 US2011159305 A1 US 2011159305A1
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peg
passivating
metal
polyethylene glycol
substituted
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Steven M. Menchen
Christina E. Inman
Meng Taing
Khai Luong
Handong Li
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Applied Biosystems LLC
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/551Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T428/00Stock material or miscellaneous articles
    • Y10T428/31504Composite [nonstructural laminate]
    • Y10T428/31678Of metal

Definitions

  • This application relates generally to surface modification techniques and, in particular, to intermediates and methods for forming passivated surfaces on oxide layers and to articles produced thereby.
  • DNA sequencing in real time requires the interrogation of individual incorporation events by polymerases, preferably on surfaces.
  • these incorporation events are identified by examination of labels on the incorporated nucleotides as individual molecules (i.e., single molecule detection).
  • the most popular labels are fluorescent dyes. Even rare non-specific adsorption of labeled nucleotides on the surfaces creates false positive signals. Therefore, binding of the substrates to the surfaces should be minimized.
  • a method of modifying a surface of a support which comprises:
  • metal reagent has a structure represented by the formula:
  • Y is a transition metal, magnesium or aluminum
  • L is oxygen, sulfur, selenium or an amine
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon;
  • n is an integer.
  • the method according to this embodiment can further comprise contacting the modified surface with a polyvalent reagent comprising a passivating moiety and a plurality of functional groups that are reactive with Y or that form complexes with Y, wherein the passivating moiety is selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon.
  • X is O, N or a methylene group
  • R 3 is polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon;
  • R 4 is N(R 2 ) 4 , or M, wherein M is Li, Na, K or Cs and wherein R 2 is an alkyl group.
  • a compound which comprises a metal complexing agent conjugated to a passivating agent, wherein:
  • the metal complexing agent is selected from the group consisting of a carboxylate, dopamine and anachelin; and wherein
  • the passivating agent is selected from the group consisting of a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon.
  • Y is a transition metal, magnesium or aluminum
  • L is oxygen, sulfur, selenium or an amine
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and
  • n is an integer.
  • a method which comprises:
  • Y is a transition metal, magnesium or aluminum
  • R 6 is a C 1 -C 5 hydrocarbon
  • L is oxygen, sulfur, selenium or an amine
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and
  • n is an integer.
  • a solid support which comprises:
  • an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin;
  • moieties on a surface of the oxide layer having a structure as set forth in formula (I) or formula (II) below:
  • X is O, N or a methylene group
  • Y is a transition metal, magnesium or aluminum
  • L is oxygen, sulfur, selenium or an amine or L is a metal complexing agent
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon;
  • R 3 is polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon;
  • oxygen atoms are bonded to the same or to different silicon, aluminum, germanium, gallium, indium or tin atoms in the oxide layer.
  • Non-limiting examples of the metal complexing agent include carboxylate, dopamine and anachelin.
  • a solid support which comprises:
  • an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin;
  • atoms of a metal selected from the group consisting of a transition metal, magnesium and aluminum bonded to the oxide layer;
  • a passivating reagent comprising a passivating moiety selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon;
  • the passivating reagent is covalently bonded to or forms a complex with one or more atoms of the metal at a plurality of locations.
  • FIG. 1A is a schematic depicting an oxide surface comprising hydroxyl groups which can be passivated according to one embodiment.
  • FIG. 1B is a schematic depicting surfaces comprising hydroxide anion groups which can be passivated according to a further embodiment.
  • FIG. 2 shows the preparation of a metal reagent which can be used to modify oxide surfaces.
  • FIG. 3 shows a modified surface resulting from the reaction of a metal reagent compound with the oxide surface.
  • FIGS. 4A , 4 B and 4 C illustrate the chemical structures of passivating agents containing phosphate groups which can be reacted with surfaces treated with a metal reagent to form passivated surfaces.
  • FIG. 5 is a schematic showing the result of the interaction of a metal reagent treated surface with a passivating agent comprising a phosphate group.
  • FIG. 6 illustrates the displacement of a labile monovalent passivating agent on a surface with a multivalent PEG passivating agent comprising a plurality of hydroxyl groups.
  • FIGS. 7A-7E are plots showing the results from confocal fluorescence imaging of coverslips with TaPEG 550 /TEG phosphate modified surfaces wherein the plots are histograms of the count rates in the pixels of a 100 ⁇ 100 ⁇ m image.
  • FIGS. 8A-8C are plots showing count rate histograms of a TaPEG 550 (no phosphate) modified surface using sulfo-Liz dCTP.
  • FIG. 9A is a correlation curve obtained from 0.1 nM Liz-dGTP on conventionally prepared 5% Pegsilane modified coverslips showing a tail at long lag times (e.g., 10 ms) indicative of sticky dyes at the surface.
  • FIGS. 9B and 9C show the correlation curves obtained from fluorescence correlation spectroscopy (FCS) of dilute solutions of rhodamine dyes and sulfoLiz dCTP diffusing above a TaPEG 550 surface modified coverslip.
  • FCS fluorescence correlation spectroscopy
  • FIG. 10A shows non-specific binding data for 1 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 10B shows non-specific binding data for a 1 nm fluorescent LIZ dye solution after 4 hours in the presence of the buffer on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 10C shows non-specific binding data for 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 10D shows non-specific binding data for a 100 nm fluorescent LIZ dye solution after 4 hours in the presence of the buffer on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 11A shows non-specific binding data for 1 nM fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 .
  • FIG. 11B shows non-specific binding data for 100 nM fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 .
  • FIG. 11C shows non-specific binding data for 1 nM fluorescent LIZ dye solution after 4 hours in the presence of a buffer on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 .
  • FIGS. 12A and 12B are photographs showing fused silica treated with TaPEG 550 then PEG 350 -triphosphate before ( FIG. 12A ) and after ( FIG. 12B ) incubation in DNA extension buffer wherein the slides were incubated with 100 nM LIZ-dATP and then rinsed with water to remove any unstuck dye.
  • a technique for forming very thin layers of aluminum or transition metal compounds (e.g., esters) onto oxide (e.g., silica) surfaces, with the object of creating a passivated surface or creating a modified surface for reaction with a functional group on the end of a passivating agent, is provided.
  • the passivating agent can be polyethylene glycol (i.e, PEG), which does not self assemble.
  • the functional group can be a phosphate or a diphosphate group with a tetrabutyl ammonium counter ion, to make the reagent compatible for use in organic solvents.
  • Transition metal esters of Ta(5+), Ti(4+), and Zr(4+) are commercially available (e.g., as methyl, ethyl, and isopropyl esters).
  • these esters with trace amounts of water to produce insoluble oxides precludes their use in an aqueous environment for coating surfaces.
  • the present inventors have found that the commercial esters of Ta(5+) and Ti(4+) can be reacted with four or more equivalents of medium molecular weight (i.e., 2 or more EO units) PEG mono-methyl ether to create a reagent with excellent solubility in most organic solvents, and with the capability of forming very thin layers of those metals on oxide surfaces such as silica surfaces, in a very predictable manner.
  • the deposition creates a passivated surface that is suitable for use in single-molecule detection schemes.
  • the modified surface can be further reacted with a phosphate or diphosphate derivative of PEG or another passivating agent to create a desired surface.
  • FIGS. 1-5 The overall scheme is summarized in FIGS. 1-5 .
  • FIG. 1A is a schematic depiction of the surface of an oxide material (e.g., silicate glass or quartz) which contains reactive hydroxyl groups. Hydroxyl groups on the surface can be converted to hydroxide anions by treatment with a base.
  • FIG. 1B is a schematic depiction of the surface of an oxide material (e.g., silicate glass or quartz) which contains reactive hydroxide anion groups. According to one embodiment, hydroxyl groups on the surface can be converted to hydroxide anions by treatment with tetrabutylammonium hydroxide. This results in the formation of the quaternary ammonium salt which can be very easily solvated in organic media.
  • an oxide material e.g., silicate glass or quartz
  • the oxide surface can be treated prior to passivation with an oxygen plasma or with tetrabutylammonium hydroxide, potassium hydroxide in methanol, hydrogen peroxide in sulfuric acid (i.e., “piranha” solution), nitric acid in sulfuric acid (i.e., “aquaregia”), hydrogen peroxide in ammonia (i.e., “RCA” solution), sulfuric acid, hydrofluoric acid, EDTA, or successive combinations of these treatments.
  • an oxygen plasma or with tetrabutylammonium hydroxide, potassium hydroxide in methanol
  • hydrogen peroxide in sulfuric acid i.e., “piranha” solution
  • nitric acid in sulfuric acid i.e., “aquaregia”
  • hydrogen peroxide in ammonia i.e., “RCA” solution
  • sulfuric acid hydrofluoric acid
  • EDTA or successive combinations of these treatments.
  • FIG. 2 illustrates the preparation of a metal reagent which can be used to modify oxide surfaces.
  • a metal reagent which can be used to modify oxide surfaces.
  • more than the number of required equivalents (i.e., m) of the Pol-LH compound can be added to get total replacement of the low molecular weight alcohols on the metal.
  • the PEG should be dry prior to addition to the metal ester. After several hours, the solution can be evacuated to remove displaced low molecular weight alcohol which aids in driving the reaction to completion.
  • FIG. 3 shows the result of the reaction of the metal reagent with the oxide surface.
  • residual “Pol” e.g., PEG
  • PEG polyethylene glycol
  • the presence of these “Pol” groups enables the use of a variety of solvents (e.g., ethyl acetate, toluene, methylene chloride, as examples) to take advantage of differing polarities, and refluxing temperatures, if either is required to optimize the speed of the deposition. It has been found that modification of the surface with the metal reagent results in a very thin layer (as demonstrated by elemental analysis at the surface by XPS) under a variety of conditions.
  • FIG. 4A shows the chemical structure of a polymeric passivating agent with a complexing agent such as a phosphate or phosphonate which can be used to treat metal-reagent modified oxide surfaces to form passivated surfaces.
  • the substituent R 3 can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • the phosphate esters can be converted to their salts (e.g., tetrabutylammonium salts) to improve solubility in a variety of organic solvents.
  • the substituent R 4 can be N(R 2 ) 4 or M wherein R 2 is an alkyl group and M is Li, Na, K or Cs and the substituent R 3 can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • the polymeric passivating agent can contain substitutions to provide all or part of the surface with a desired functionality.
  • the R 3 substituent of the phosphate ester shown in FIGS. 4A-4C may include biotin moieties.
  • the metal reagent treated surface can be contacted with a composition comprising a mixture of passivating agents having different polymeric elements to provide a surface optimized for a particular end-use application.
  • the passivating formulation may contain a mixture of PEG phosphates or diphosphates with fluorocarbon phosphates or diphosphates in order to obtain desired surface properties. The desired surface properties can be determined empirically, through experimentation. In FIGS.
  • X can be O, N or a methylene group and the R 3 substituent can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • each R 4 substituent can be N(R 2 ) 4 , or M, wherein M is Li, Na, K or Cs and wherein R 2 is an alkyl group.
  • FIG. 5 shows a surface formed by treating an oxide surface with a passivating agent comprising a phosphate group wherein the oxide surface has been previously treated with a metal reagent.
  • the R 3 substituent can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • Y can be a transition metal, magnesium or aluminum and Z can be O, N or a methylene group.
  • metal reagents described herein enables controlled formation of very thin layers of metal precursor on a variety of oxide surfaces.
  • the surfaces can be formed under very mild conditions.
  • the metal reagents are also easy to make and require no purification prior to use.
  • the use of a metal reagent pre-layer and a phosphate functional passivating agent enables the use of readily available phosphate functionality for the deposition of the desired surface.
  • the phosphates can be readily synthesized from phosphorous oxychloride and the alcohol versions of the passivating agents.
  • the synthesized phosphates can be converted to derivatives (e.g., quaternary ammonium derivatives) that are readily soluble in a variety of solvents, and that still maintain their reactivity with the metal reagent modified surfaces. Reaction of the phosphate with the surface does not require extensive heating or self-assembly of the tailed phosphates on the surface.
  • metal reagents and phosphate functional passivating agents also enables the creation of a variety of surface types and properties by formulating phosphate esters with different polymeric groups.
  • phosphates of hydrocarbon, fluorocarbon, and PEG alcohols could be mixed together prior to contact with the metal-reagent treated oxide surface to obtain a modified surface with desirable properties for a specific application.
  • a phosphate ester containing a specific functionality e.g., biotin
  • streptavidin e.g., streptavidin
  • the surface of a support treated with a metal reagent as described above can be subsequently treated with a multivalent passivating agent comprising a passivating moiety and a plurality of functional groups capable of forming covalent bonds or forming complexes with metal atoms on the surface.
  • the multivalent passivating agent can displace the monovalent passivating agent on the transition metal oxide surface while maintaining the transition metal layer between the oxide surface and the passivating agent.
  • FIG. 6 A schematic depiction of the displacement of the monovalent passivating agent with a multivalent passivating agent is illustrated in FIG. 6 . In this manner, the multivalent passivating agent can impart substantial hydrolytic stability to the passivated surface.
  • the passivating moiety on the multivalent passivating agent can be a moiety selected from the group consisting of: a substituted polyethylene glycol; an unsubstituted polyethylene glycol; a hydrocarbon; a substituted hydrocarbon; a fluorocarbon and a substituted fluorocarbon.
  • the plurality of functional groups that are reactive with the metal atoms on the surface or that form complexes with the metal atoms on the surface can be hydroxyl groups, amine groups, phosphate groups, phosphonate groups, thiol groups, alkylphosphate groups, carboxyl groups or combinations thereof.
  • R is H, alkyl, aryl or a functional group; L is a linker group or a covalent bond; each W and Y are independently O, NH, S or phosphonate; each X and Z are independently H, phosphate, alkylcarboxy or alkylphosphate; n is 3 to 100; o is 1-8; p is 0-8; and q is 1-3.
  • the multivalent passivating agent comprises a unsubstituted polyethylene glycol as the passivating moiety. Although an unsubstituted polyethylene glycol passivating moiety is depicted, other passivating moieties as described above can also be employed.
  • a method of synthesizing a multivalent passivating agent comprising a polyethylene glycol passivating moiety and a plurality of hydroxyl functional groups is shown below.
  • an amino-functional compound comprising a polyethylene glycol moiety can be reacted with 3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-one at 80° C. to form the multivalent passivating agent.
  • the variables in the general formula above are defined as follows: R is methyl; n is 11, s is 1, q is 1, M is C ⁇ O, N is —NH—C—C—NH—(C ⁇ O)—, o is 4, W and Y are O, X and Z are H, and p is 1.
  • linker L in the above formula can be a linker group.
  • exemplary linker groups include, but are not limited to, a group represented by the formula: —(C) s —(C ⁇ O)-Q-R′-Q-(C ⁇ O)— wherein each Q is independently —O— or —NH—, R′ is an aliphatic or aromatic group and s is an integer of 0 to 10.
  • n is a positive integer.
  • n can be 3 to 100.
  • the polyvalent reagent can be represented by the following formula:
  • n is a positive integer.
  • n can be 3 to 100.
  • a solid support having a surface treated with a multivalent passivating agent as set forth above is also provided.
  • a solid support which comprises: an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin; atoms of a metal selected from the group consisting of a transition metal, magnesium and aluminum bonded to the oxide layer; and a passivating reagent comprising a passivating moiety selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon; wherein the passivating reagent is covalently bonded to or forms a complex with one or more atoms of the metal at a plurality of locations.
  • modification of a surface with metal esters enables very controlled, very thin layer deposition of a metal precursor to a variety of oxide surfaces under very mild conditions.
  • the metal esters are very easy to make, and require no purification prior to use.
  • the use of a combination of metal ester deposition followed by multivalent passivating agent deposition can enable the creation of a variety of surfaces types and properties.
  • the surface can be treated with a multivalent passivating agent having different functional groups during the final surface modification.
  • the multivalent passivating agent can be readily synthesized using known chemical synthesis techniques.
  • An exemplary synthesis technique is described above for a multivalent passivating agent comprising polyethylene glycol as a passivating moiety and a plurality of hydroxyl functional groups capable of bonding to metal atoms on a support surface.
  • a method for synthesizing a multivalent passivating agent comprising a polyethylene glycol passivating moiety and a plurality of phosphate moieties is described below.
  • the phosphate moieties are capable of forming complexes with metal atoms on a support surface.
  • a first step of the method comprises the synthesis of a Methyl PEG 2000 -Acetic Acid intermediate.
  • This intermediate can be synthesized by the procedure set forth below.
  • Methyl-PEG 2000 alcohol (100 gram, av. 0.05 mole) was dissolved in 100 ml anhydrous dichloromethane (DCM) with NaOH (6 g, 0.15 moles) added and chilled on ice under nitrogen.
  • DCM anhydrous dichloromethane
  • the Methyl PEG2K-Acetic Acid intermediate reaction product can then be reacted with 2-amino-2-(hydroxymethyl)propane-1,3-diol as set forth below to form a second intermediate comprising a plurality of hydroxyl groups (i.e., an mPEG2K Acetamido-Tris-Alcohol).
  • Methyl PEG 2000 -acetic acid (12 gram, av. 0.006 mole) was co-evaporated with anhydrous toluene and acetonitrile, respectively three times (30 ml each) and redissolved in dry dichloromethane (10 ml).
  • Triethylamine (TEA) (0.75 ml, 0.006 mole) was added under dry nitrogen.
  • DSC Disuccinimidyl carbonate
  • R f value of the product was about 0.5 in 10% MeOH-DCM on a silica TLC plate.
  • the mPEG 2000 Acetamido-Tris-Alcohol intermediate can then be reacted with bis(2-cyanoethyl)diisopropylphosphoramidite as set forth below to form the multivalent passivating agent.
  • R f of the product was 0.55 in 10% MeOH-DCM on a silica TLC plate.
  • methyl PEG-350 acetamido-Tris-Triphosphate was synthesized following the same procedure for the synthesis of methyl PEG-2K acetamido-Tris-Triphosphate.
  • XPS data are shown in the table below for a tantalum PEG-ester modified silicon dioxide surface after performing the following manipulations on the surface:
  • XPS data are shown in the table below for a titanium PEG-ester modified silicon dioxide surface after performing the following manipulations on the surface:
  • FIGS. 7A-7E are plots showing the results from confocal fluorescence imaging of coverslips with TaPEG 550 /TEG phosphate modified surfaces.
  • the plots are histograms of the count rates in the pixels of a 100 ⁇ 100 ⁇ m image.
  • FIG. 7A shows that, before any dye is added, the background fluorescence signal peaks around 2.5 kHz.
  • FIGS. 7B and 7C show that when a 1 nM A488 solution is added, the count rate histogram peak shifts to 70 kHz and drops again back to signal levels comparable to the background when the dye is removed. This is repeated with a 1 nM R110 solution in the same 100 ⁇ 100 ⁇ m region as shown in FIG. 7D .
  • the results show that the fluorescence signals return to background levels when R110 is removed from the coverslip ( FIG. 7E ).
  • FIGS. 8A-8C are plots showing similar count rate histograms of a TaPEG 550 (no phosphate) modified surface using sulfo-Liz dCTP which demonstrate that the modified surface prevents sulfo-Liz dCTP from nonspecifically binding to the coverslip. This is observed even when a high concentration sulfo-Liz dCTP solution is added onto the surface ( FIG. 8B ) and removed ( FIG. 8C ). Again, these results demonstrate that sulfo-Liz dCTP does not non-specifically bind to the modified surface.
  • FIGS. 9A-9C illustrate a comparison of the correlation curves obtained from fluorescence correlation spectroscopy (FCS) of dilute solutions of the above rhodamine dyes and the sulfoLiz dCTP in contact with TaPEG 550 and silane PEG surface modified coverslips.
  • FCS fluorescence correlation spectroscopy
  • the decay rate of the correlation curves provide information on the rate of diffusion of the free dyes in solution. Hence, if a dye sticks to the surface, the correlation curve will decay more slowly.
  • FIG. 9A is an example of a correlation curve exhibiting a tail at long lag times (e.g., 10 ms) indicative of sticky dyes at the surface.
  • the data for FIG. 9A was obtained from 0.1 nM Liz-dGTP on conventionally prepared 5% PEGsilane modified coverslips wherein the PEGsilane was deposited from refluxing toluene.
  • FIG. 9B is a plot showing the correlation curves from dilute solutions of A488 and R110 diffusing above the modified surface. This plot also shows the correlation curve for freely diffusing A488 above a known non-sticky surface for comparison. FCS experiments were also done using dilute solutions of sulfo-Liz dCTP, the results of which are shown in FIG. 9C . This plot shows the results from three different FCS measurements at different positions on the modified coverslip. Together, these FCS results and the fluorescence imaging results above indicate that the TaPEG 550 modified surfaces on fused silica substrate do not non-specifically bind A488, R110, and sulfo-Liz dCTP.
  • a quartz surface was treated with ethanolic Ta(monomethyl-PEG 550 ) 5 to form a modified surface.
  • the modified surface was then treated with an ethanolic solution of a multivalent passivating agent having the following structure:
  • FIG. 10A shows non-specific binding data for 1 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 10B shows non-specific binding data for a 1 nm fluorescent LIZ dye solution treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 after it had been incubated in a Tris buffer solution for 4 hours.
  • FIG. 10A shows non-specific binding data for 1 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 after it had been incubated in a Tris buffer solution for 4 hours.
  • FIG. 10C shows non-specific binding data for 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 .
  • FIG. 10D shows non-specific binding data for a 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG 550 ) 5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG 550 after the surface had been incubated in a Tris buffer solution for 4 hours.
  • treatment with the multivalent passivating agent yields a surface that prevents non-specific adsorption of a 100 nM aqueous solution of Liz fluorescent dye even after 4 hours in the presence of a buffer.
  • the integrity of the Ta surface after treatment with the multivalent passivating agent i.e., poly(hydroxyl)-monomethyl-PEG
  • the integrity of the Ta surface after treatment with the multivalent passivating agent is demonstrated by XPS of the modified quartz surfaces before and after poly(hydroxyl)-monomethyl-PEG modification, shown below.
  • FIGS. 12A and 12B show fused silica surfaces treated with TaPEG 550 then PEG 350 triphosphate before ( FIG. 12A ) and after ( FIG. 12B ) incubation in DNA extension buffer.
  • the PEG 350 triphosphate has a chemical structure as set forth below:
  • n has a value such that the polyethylene glycol moiety has an average molecular weight of about 350.
  • the treated surfaces were incubated with 100 nM LIZ-dATP and then rinsed with water to remove any dye that was not stuck to the surface. As can be seen from FIG. 12B , 100 nM LIZ-dATP exhibits minimal sticking to the modified surface even after the treated slide has been incubated with DNA extension buffer.

Abstract

Intermediates and methods for forming passivated surfaces on oxide layers and articles produced thereby are described. Hydroxyl or hydroxide groups on the oxide surfaces are reacted with a metal reagent of the formula Y(L-Pol)m, where Y is a transition metal, magnesium or aluminum, L is oxygen, sulfur, selenium or an amine, and “Pol” represents a passivating agent such as a polyethylene glycol, a hydrocarbon, or a fluorocarbon. The resulting modified surface can be further reacted with a passivating agent having a phosphate functional group or a polyvalent reagent comprising a passivating moiety and a plurality of functional groups that are reactive with or that form complexes with Y. The passivating agent can also include a functional group such as biotin to provide surfaces with a desired functionality. The passivated surfaces exhibit minimal binding to bio-molecules and can be used in single-molecule detection schemes.

Description

  • This application is a divisional of U.S. patent application Ser. No. 11/943,851, filed on Nov. 21, 2007, which claims the benefit of Provisional U.S. Patent Application Ser. No. 60/860,215, filed on Nov. 21, 2006, and Provisional U.S. Patent Application Ser. No. 60/860,480, filed on Nov. 22, 2006. Each of the above applications is incorporated by reference herein in its entirety.
  • The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described herein in any way.
  • BACKGROUND
  • 1. Technical Field
  • This application relates generally to surface modification techniques and, in particular, to intermediates and methods for forming passivated surfaces on oxide layers and to articles produced thereby.
  • 2. Background of the Technology
  • DNA sequencing in real time requires the interrogation of individual incorporation events by polymerases, preferably on surfaces. Generally, these incorporation events are identified by examination of labels on the incorporated nucleotides as individual molecules (i.e., single molecule detection). The most popular labels are fluorescent dyes. Even rare non-specific adsorption of labeled nucleotides on the surfaces creates false positive signals. Therefore, binding of the substrates to the surfaces should be minimized.
  • There still exists a need for improved passivation techniques which would allow for the formation of thin passivation layers and which would result in surfaces exhibiting minimal absorption of biomolecules.
  • SUMMARY
  • According to a first embodiment, a method of modifying a surface of a support is provided which comprises:
  • contacting the surface with a metal reagent, wherein the surface comprises hydroxyl and/or oxide anion groups; and
  • allowing the metal reagent to react with hydroxyl and/or oxide anion groups on the surface to form a modified surface;
  • wherein the metal reagent has a structure represented by the formula:

  • Y(L-Pol)m
  • wherein:
  • Y is a transition metal, magnesium or aluminum;
  • L is oxygen, sulfur, selenium or an amine;
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon; and
  • m is an integer.
  • The method according to this embodiment can further comprise contacting the modified surface with a polyvalent reagent comprising a passivating moiety and a plurality of functional groups that are reactive with Y or that form complexes with Y, wherein the passivating moiety is selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon.
  • According to a second embodiment, a compound is provided which is represented by the formula (I), formula (II), or formula (III) below:
  • Figure US20110159305A1-20110630-C00001
  • wherein:
  • X is O, N or a methylene group;
  • R3 is polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon; and
  • R4 is N(R2)4, or M, wherein M is Li, Na, K or Cs and wherein R2 is an alkyl group.
  • According to a third embodiment, a compound is provided which comprises a metal complexing agent conjugated to a passivating agent, wherein:
  • the metal complexing agent is selected from the group consisting of a carboxylate, dopamine and anachelin; and wherein
  • the passivating agent is selected from the group consisting of a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon.
  • According to a fourth embodiment, a compound is provided which is represented by the formula:

  • Y(L-Pol)m
  • wherein:
  • Y is a transition metal, magnesium or aluminum,
  • L is oxygen, sulfur, selenium or an amine,
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and
  • m is an integer.
  • According to a fifth embodiment, a method is provided which comprises:
  • reacting a first compound of the formula:

  • Y(OR6)m
  • with one or more second compounds each of the formula:

  • Pol-LH
  • to form a metal ester compound of the formula

  • Y(L-Pol)m
  • wherein:
  • Y is a transition metal, magnesium or aluminum,
  • R6 is a C1-C5 hydrocarbon,
  • L is oxygen, sulfur, selenium or an amine,
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and
  • m is an integer.
  • According to a sixth embodiment, a solid support is provided which comprises:
  • an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin; and
  • moieties on a surface of the oxide layer, the moieties having a structure as set forth in formula (I) or formula (II) below:
  • Figure US20110159305A1-20110630-C00002
  • wherein:
  • X is O, N or a methylene group;
  • Y is a transition metal, magnesium or aluminum;
  • L is oxygen, sulfur, selenium or an amine or L is a metal complexing agent;
  • each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon; and
  • R3 is polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon; and
  • wherein the oxygen atoms are bonded to the same or to different silicon, aluminum, germanium, gallium, indium or tin atoms in the oxide layer.
  • Non-limiting examples of the metal complexing agent include carboxylate, dopamine and anachelin.
  • According to a seventh embodiment, a solid support is provided which comprises:
  • an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin;
  • atoms of a metal selected from the group consisting of a transition metal, magnesium and aluminum bonded to the oxide layer; and
  • a passivating reagent comprising a passivating moiety selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon;
  • wherein the passivating reagent is covalently bonded to or forms a complex with one or more atoms of the metal at a plurality of locations.
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • The skilled artisan will understand that the drawings, described below, are for illustration purposes only. The drawings are not intended to limit the scope of the present teachings in any way.
  • FIG. 1A is a schematic depicting an oxide surface comprising hydroxyl groups which can be passivated according to one embodiment.
  • FIG. 1B is a schematic depicting surfaces comprising hydroxide anion groups which can be passivated according to a further embodiment.
  • FIG. 2 shows the preparation of a metal reagent which can be used to modify oxide surfaces.
  • FIG. 3 shows a modified surface resulting from the reaction of a metal reagent compound with the oxide surface.
  • FIGS. 4A, 4B and 4C illustrate the chemical structures of passivating agents containing phosphate groups which can be reacted with surfaces treated with a metal reagent to form passivated surfaces.
  • FIG. 5 is a schematic showing the result of the interaction of a metal reagent treated surface with a passivating agent comprising a phosphate group.
  • FIG. 6 illustrates the displacement of a labile monovalent passivating agent on a surface with a multivalent PEG passivating agent comprising a plurality of hydroxyl groups.
  • FIGS. 7A-7E are plots showing the results from confocal fluorescence imaging of coverslips with TaPEG550/TEG phosphate modified surfaces wherein the plots are histograms of the count rates in the pixels of a 100×100 μm image.
  • FIGS. 8A-8C are plots showing count rate histograms of a TaPEG550 (no phosphate) modified surface using sulfo-Liz dCTP.
  • FIG. 9A is a correlation curve obtained from 0.1 nM Liz-dGTP on conventionally prepared 5% Pegsilane modified coverslips showing a tail at long lag times (e.g., 10 ms) indicative of sticky dyes at the surface.
  • FIGS. 9B and 9C show the correlation curves obtained from fluorescence correlation spectroscopy (FCS) of dilute solutions of rhodamine dyes and sulfoLiz dCTP diffusing above a TaPEG550 surface modified coverslip.
  • FIG. 10A shows non-specific binding data for 1 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550.
  • FIG. 10B shows non-specific binding data for a 1 nm fluorescent LIZ dye solution after 4 hours in the presence of the buffer on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550.
  • FIG. 10C shows non-specific binding data for 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550.
  • FIG. 10D shows non-specific binding data for a 100 nm fluorescent LIZ dye solution after 4 hours in the presence of the buffer on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550.
  • FIG. 11A shows non-specific binding data for 1 nM fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5.
  • FIG. 11B shows non-specific binding data for 100 nM fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5.
  • FIG. 11C shows non-specific binding data for 1 nM fluorescent LIZ dye solution after 4 hours in the presence of a buffer on a quartz surface treated with Ta(monomethyl-PEG550)5.
  • FIGS. 12A and 12B are photographs showing fused silica treated with TaPEG550 then PEG350-triphosphate before (FIG. 12A) and after (FIG. 12B) incubation in DNA extension buffer wherein the slides were incubated with 100 nM LIZ-dATP and then rinsed with water to remove any unstuck dye.
  • DETAILED DESCRIPTION
  • The use of “or” herein means “and/or” unless stated otherwise or where the use of “and/or” is clearly inappropriate. The use of “a” herein means “one or more” unless stated otherwise or where the use of “one or more” is clearly inappropriate. The use of “comprise,” “comprises,” “comprising,” “include,” “includes,” and “including” are interchangeable and not intended to be limiting. Furthermore, where the description of one or more embodiments uses the term “comprising,” those skilled in the art would understand that, in some specific instances, the embodiment or embodiments can be alternatively described using the language “consisting essentially of” and/or “consisting of.” It should also be understood that in some embodiments the order of steps or order for performing certain actions is immaterial so long as the present teachings remain operable. Moreover, in some embodiments two or more steps or actions can be conducted simultaneously.
  • A technique for forming very thin layers of aluminum or transition metal compounds (e.g., esters) onto oxide (e.g., silica) surfaces, with the object of creating a passivated surface or creating a modified surface for reaction with a functional group on the end of a passivating agent, is provided. The passivating agent can be polyethylene glycol (i.e, PEG), which does not self assemble. The functional group can be a phosphate or a diphosphate group with a tetrabutyl ammonium counter ion, to make the reagent compatible for use in organic solvents.
  • Transition metal esters of Ta(5+), Ti(4+), and Zr(4+) are commercially available (e.g., as methyl, ethyl, and isopropyl esters). However, the reactivity of these esters with trace amounts of water to produce insoluble oxides precludes their use in an aqueous environment for coating surfaces. In addition, their limited solubility in organic solvents, especially the commercially available esters of Ti(4+), precludes the use of these reagents in many organic solvents.
  • The present inventors have found that the commercial esters of Ta(5+) and Ti(4+) can be reacted with four or more equivalents of medium molecular weight (i.e., 2 or more EO units) PEG mono-methyl ether to create a reagent with excellent solubility in most organic solvents, and with the capability of forming very thin layers of those metals on oxide surfaces such as silica surfaces, in a very predictable manner. In some cases, the deposition creates a passivated surface that is suitable for use in single-molecule detection schemes. In other cases, the modified surface can be further reacted with a phosphate or diphosphate derivative of PEG or another passivating agent to create a desired surface.
  • The overall scheme is summarized in FIGS. 1-5.
  • FIG. 1A is a schematic depiction of the surface of an oxide material (e.g., silicate glass or quartz) which contains reactive hydroxyl groups. Hydroxyl groups on the surface can be converted to hydroxide anions by treatment with a base. FIG. 1B is a schematic depiction of the surface of an oxide material (e.g., silicate glass or quartz) which contains reactive hydroxide anion groups. According to one embodiment, hydroxyl groups on the surface can be converted to hydroxide anions by treatment with tetrabutylammonium hydroxide. This results in the formation of the quaternary ammonium salt which can be very easily solvated in organic media.
  • The oxide surface can be treated prior to passivation with an oxygen plasma or with tetrabutylammonium hydroxide, potassium hydroxide in methanol, hydrogen peroxide in sulfuric acid (i.e., “piranha” solution), nitric acid in sulfuric acid (i.e., “aquaregia”), hydrogen peroxide in ammonia (i.e., “RCA” solution), sulfuric acid, hydrofluoric acid, EDTA, or successive combinations of these treatments.
  • FIG. 2 illustrates the preparation of a metal reagent which can be used to modify oxide surfaces. In order to synthesize the reagent, more than the number of required equivalents (i.e., m) of the Pol-LH compound can be added to get total replacement of the low molecular weight alcohols on the metal. The PEG should be dry prior to addition to the metal ester. After several hours, the solution can be evacuated to remove displaced low molecular weight alcohol which aids in driving the reaction to completion.
  • FIG. 3 shows the result of the reaction of the metal reagent with the oxide surface. In principle, residual “Pol” (e.g., PEG) groups are left on the un-reacted sites of the metal-ester, which are used later for conversion to the insoluble metal-partner pair. The presence of these “Pol” groups enables the use of a variety of solvents (e.g., ethyl acetate, toluene, methylene chloride, as examples) to take advantage of differing polarities, and refluxing temperatures, if either is required to optimize the speed of the deposition. It has been found that modification of the surface with the metal reagent results in a very thin layer (as demonstrated by elemental analysis at the surface by XPS) under a variety of conditions.
  • In some cases, the treatment of the oxide surface with the metal reagent yields the passivated surface. In other cases, the modified surface formed by treatment with the metal reagent is an intermediate to the formation of a passivated surface with an additional passivating step. FIG. 4A shows the chemical structure of a polymeric passivating agent with a complexing agent such as a phosphate or phosphonate which can be used to treat metal-reagent modified oxide surfaces to form passivated surfaces. In FIG. 4A, the substituent R3 can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • As indicated in FIGS. 4B and 4C, the phosphate esters can be converted to their salts (e.g., tetrabutylammonium salts) to improve solubility in a variety of organic solvents. In FIGS. 4B and 4C, the substituent R4 can be N(R2)4 or M wherein R2 is an alkyl group and M is Li, Na, K or Cs and the substituent R3 can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon.
  • In addition, the polymeric passivating agent can contain substitutions to provide all or part of the surface with a desired functionality. For example, the R3 substituent of the phosphate ester shown in FIGS. 4A-4C may include biotin moieties. In addition, the metal reagent treated surface can be contacted with a composition comprising a mixture of passivating agents having different polymeric elements to provide a surface optimized for a particular end-use application. For example, the passivating formulation may contain a mixture of PEG phosphates or diphosphates with fluorocarbon phosphates or diphosphates in order to obtain desired surface properties. The desired surface properties can be determined empirically, through experimentation. In FIGS. 4A-4C, X can be O, N or a methylene group and the R3 substituent can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon. Also in FIGS. 4B and 4C, each R4 substituent can be N(R2)4, or M, wherein M is Li, Na, K or Cs and wherein R2 is an alkyl group.
  • FIG. 5 shows a surface formed by treating an oxide surface with a passivating agent comprising a phosphate group wherein the oxide surface has been previously treated with a metal reagent. As can be seen from FIG. 5, this technique results in a thin-layer of the passivating agent on the oxide surface. In FIG. 5, the R3 substituent can be polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon. Also in FIG. 5, Y can be a transition metal, magnesium or aluminum and Z can be O, N or a methylene group.
  • The use of the metal reagents described herein enables controlled formation of very thin layers of metal precursor on a variety of oxide surfaces. In addition, the surfaces can be formed under very mild conditions. The metal reagents are also easy to make and require no purification prior to use.
  • The use of a metal reagent pre-layer and a phosphate functional passivating agent enables the use of readily available phosphate functionality for the deposition of the desired surface. The phosphates can be readily synthesized from phosphorous oxychloride and the alcohol versions of the passivating agents. The synthesized phosphates can be converted to derivatives (e.g., quaternary ammonium derivatives) that are readily soluble in a variety of solvents, and that still maintain their reactivity with the metal reagent modified surfaces. Reaction of the phosphate with the surface does not require extensive heating or self-assembly of the tailed phosphates on the surface.
  • The use of metal reagents and phosphate functional passivating agents also enables the creation of a variety of surface types and properties by formulating phosphate esters with different polymeric groups. For example, phosphates of hydrocarbon, fluorocarbon, and PEG alcohols could be mixed together prior to contact with the metal-reagent treated oxide surface to obtain a modified surface with desirable properties for a specific application. In addition, a phosphate ester containing a specific functionality (e.g., biotin) could be used to obtain a surface that would bind another molecule (e.g., streptavidin).
  • Treatment of Metal Reagent Modified Surfaces with a Multivalent Passivating Agent
  • According to some embodiments, the surface of a support treated with a metal reagent as described above can be subsequently treated with a multivalent passivating agent comprising a passivating moiety and a plurality of functional groups capable of forming covalent bonds or forming complexes with metal atoms on the surface. For example, the multivalent passivating agent can displace the monovalent passivating agent on the transition metal oxide surface while maintaining the transition metal layer between the oxide surface and the passivating agent. A schematic depiction of the displacement of the monovalent passivating agent with a multivalent passivating agent is illustrated in FIG. 6. In this manner, the multivalent passivating agent can impart substantial hydrolytic stability to the passivated surface.
  • The passivating moiety on the multivalent passivating agent can be a moiety selected from the group consisting of: a substituted polyethylene glycol; an unsubstituted polyethylene glycol; a hydrocarbon; a substituted hydrocarbon; a fluorocarbon and a substituted fluorocarbon.
  • The plurality of functional groups that are reactive with the metal atoms on the surface or that form complexes with the metal atoms on the surface can be hydroxyl groups, amine groups, phosphate groups, phosphonate groups, thiol groups, alkylphosphate groups, carboxyl groups or combinations thereof.
  • A general formula for a multivalent passivating agent is shown below.
  • Figure US20110159305A1-20110630-C00003
  • wherein:
  • R is H, alkyl, aryl or a functional group; L is a linker group or a covalent bond; each W and Y are independently O, NH, S or phosphonate; each X and Z are independently H, phosphate, alkylcarboxy or alkylphosphate; n is 3 to 100; o is 1-8; p is 0-8; and q is 1-3. As shown in the above formula, the multivalent passivating agent comprises a unsubstituted polyethylene glycol as the passivating moiety. Although an unsubstituted polyethylene glycol passivating moiety is depicted, other passivating moieties as described above can also be employed.
  • A method of synthesizing a multivalent passivating agent comprising a polyethylene glycol passivating moiety and a plurality of hydroxyl functional groups is shown below.
  • Figure US20110159305A1-20110630-C00004
  • As shown in the above reaction scheme, an amino-functional compound comprising a polyethylene glycol moiety can be reacted with 3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-one at 80° C. to form the multivalent passivating agent. For this multivalent passivating agent, the variables in the general formula above are defined as follows: R is methyl; n is 11, s is 1, q is 1, M is C═O, N is —NH—C—C—NH—(C═O)—, o is 4, W and Y are O, X and Z are H, and p is 1.
  • As set forth above, the linker L in the above formula can be a linker group. Exemplary linker groups include, but are not limited to, a group represented by the formula: —(C)s—(C═O)-Q-R′-Q-(C═O)— wherein each Q is independently —O— or —NH—, R′ is an aliphatic or aromatic group and s is an integer of 0 to 10.
  • An exemplary polyvalent reagent corresponding to the above general formula is represented by the following formula:
  • Figure US20110159305A1-20110630-C00005
  • wherein n is a positive integer. For example, n can be 3 to 100.
  • According to some embodiments, the polyvalent reagent can be represented by the following formula:
  • Figure US20110159305A1-20110630-C00006
  • wherein n is a positive integer. For example, n can be 3 to 100.
  • A solid support having a surface treated with a multivalent passivating agent as set forth above is also provided. According to some embodiments, a solid support is provided which comprises: an oxide layer comprising an oxide of silicon, aluminum, germanium, gallium, indium, magnesium or tin; atoms of a metal selected from the group consisting of a transition metal, magnesium and aluminum bonded to the oxide layer; and a passivating reagent comprising a passivating moiety selected from the group consisting of a substituted polyethylene glycol, an unsubstituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon and a substituted fluorocarbon; wherein the passivating reagent is covalently bonded to or forms a complex with one or more atoms of the metal at a plurality of locations.
  • As set forth above, modification of a surface with metal esters enables very controlled, very thin layer deposition of a metal precursor to a variety of oxide surfaces under very mild conditions. The metal esters are very easy to make, and require no purification prior to use. The use of a combination of metal ester deposition followed by multivalent passivating agent deposition can enable the creation of a variety of surfaces types and properties. For example, the surface can be treated with a multivalent passivating agent having different functional groups during the final surface modification.
  • Synthesis of Multivalent Passivating Agents Comprising Phosphate Moieties
  • The multivalent passivating agent can be readily synthesized using known chemical synthesis techniques. An exemplary synthesis technique is described above for a multivalent passivating agent comprising polyethylene glycol as a passivating moiety and a plurality of hydroxyl functional groups capable of bonding to metal atoms on a support surface. A method for synthesizing a multivalent passivating agent comprising a polyethylene glycol passivating moiety and a plurality of phosphate moieties is described below. The phosphate moieties are capable of forming complexes with metal atoms on a support surface.
  • Synthesis and Purification of Methyl PEG2000-Acetic Acid
  • A first step of the method comprises the synthesis of a Methyl PEG2000-Acetic Acid intermediate. This intermediate can be synthesized by the procedure set forth below.
  • Figure US20110159305A1-20110630-C00007
  • This procedure is described below.
  • 1) Methyl-PEG2000 alcohol (100 gram, av. 0.05 mole) was dissolved in 100 ml anhydrous dichloromethane (DCM) with NaOH (6 g, 0.15 moles) added and chilled on ice under nitrogen.
  • 2) Bromoacetic t-butyl ester (30 g, 0.15 moles) was added portionwise with stirring. The final mixture was agitated (stirring became difficult due to formation of NaBr solid) at room temperature for 24 hours.
  • 3) The NaBr precipitate was removed by centrifugation or decanting. Remove most of DCM on a rotavap. Add hexane to extract out unreacted Bromoacetic t-butyl ester. The semi-solid after decanting hexane was adjusted to pH 1.0 with HCl (conc.) on ice and stirred overnight at room temperature.
  • 4) The product was extracted with DCM three times (total 500 ml), washed with brine, and dried with anhydrous NaSO4. Remove most of the DCM on a Rotavap. Add to cold hexane dropwise with stirring to precipitate the product.
  • 5) A white solid product (85 grams) was collected, washed with hexane, dried under vacuum, and characterized by mass spectrometry (MS) both positive and negative mode with Matrix Assisted Laser Desorption Ionization (MALDI). The yield was 83%.
  • Synthesis and Column Purification of mPEG2K Acetamido-Tris-Alcohol
  • The Methyl PEG2K-Acetic Acid intermediate reaction product can then be reacted with 2-amino-2-(hydroxymethyl)propane-1,3-diol as set forth below to form a second intermediate comprising a plurality of hydroxyl groups (i.e., an mPEG2K Acetamido-Tris-Alcohol).
  • Figure US20110159305A1-20110630-C00008
  • This procedure is described below.
  • 1) Methyl PEG2000-acetic acid (12 gram, av. 0.006 mole) was co-evaporated with anhydrous toluene and acetonitrile, respectively three times (30 ml each) and redissolved in dry dichloromethane (10 ml). Triethylamine (TEA) (0.75 ml, 0.006 mole) was added under dry nitrogen. Disuccinimidyl carbonate (DSC) (3 gram, 0.012 moles) was added portionwise under nitrogen with stifling. The reaction was left at room temperature for 4 hours and the solvent was removed on a rotavap. Anhydrous dioxane (10 ml) was added to dissolve the residue, and the solution added dropwise to a saturated trishydroxymethylaminomethane (Tris base) aqueous solution with vigorous stirring. The reaction was left for two hours at room temperature and then extracted with DCM (total 100 ml) three times. The DCM phase was dried with sodium sulfate and an oil crude was obtained after removal of solvent.
  • 2) The residue was purified on a silica gel (100 ml) column packaged in 5% MeOH-DCM and eluted with 5% MeOH-DCM. Fraction 12-20 were identified as the desired product (9.5 gram) by MALDI-MS and proton nuclear magnetic resonance spectroscopy (HNMR). Yield was 76%.
  • 3) Rf value of the product was about 0.5 in 10% MeOH-DCM on a silica TLC plate.
  • Synthesis and Purification of Methyl PEG-2K Acetamido-Tris-Triphosphate
  • The mPEG2000 Acetamido-Tris-Alcohol intermediate can then be reacted with bis(2-cyanoethyl)diisopropylphosphoramidite as set forth below to form the multivalent passivating agent.
  • Figure US20110159305A1-20110630-C00009
    Figure US20110159305A1-20110630-C00010
  • This procedure is described below.
  • 1) mPEG2K acetamido-Tris-alcohol (2.2 gram, 1 mmole) was coevaporated with anhydrous toluene and acetonitrile, respectively, three times (30 ml each). Bis(2-cyanoethyl)diisopropylphosphoramidite (FW=271.6, 1.22 g, 4.5 mmoles) and tetrazole-acetonitrile solution (15 ml, 0.45M, 6.75 mmoles) were added under nitrogen. The reaction mixture was stirred at room temperature for 3 hours. Tert-Butyl hydroperoxide (70% aqueous, 6 ml, 45 mmoles) was added and the mixture was stirred at room temperature for 2 hours. The solvent was removed on a rotavap. The residue was extracted with DCM three times and washed with sodium bicarbonate buffer (pH 9) and dried over sodium sulfate. The residue was silica gel column purified with eluent 10% methanol in DCM. Fractions 15-20 were identified as the desired product by MS and HNMR.
  • 2) Rf of the product was 0.55 in 10% MeOH-DCM on a silica TLC plate.
  • 3) The purified phosphonate ester was placed in NH3.H2O (5 ml) and incubated overnight at room temperature. The ammonia was removed under vacuum, re-dissolved in water and purified on Sephadex DEAE A-25 column (equilibrated with 2M potassium bicarbonate and washed thoroughly with water). The phosphate was eluted with 2M triethylamine acetate buffer. The buffer was removed by repeatedly co-evaporating with water under vacuum and the product (1.7 gram) was characterized by MALDI-MS, proton NMR and phosphor NMR. Yield was 70%.
  • Synthesis and Purification of Methyl PEG-350 Acetamido-Tris-Triphosphate
  • Using the starting material methyl PEG 350 alcohol with an average molecular weight 350, methyl PEG-350 acetamido-Tris-Triphosphate was synthesized following the same procedure for the synthesis of methyl PEG-2K acetamido-Tris-Triphosphate.
  • EXPERIMENTAL
  • Aspects of the present teachings may be further understood in light of the following examples, which should not be construed as limiting the scope of the present teachings in any way.
  • Surfaces Treated with Metal Reagents and TEG Phosphate
  • XPS data are shown in the table below for a tantalum PEG-ester modified silicon dioxide surface after performing the following manipulations on the surface:
  • treatment of the quartz cover-slip for 16 hr. at 77° C. temperature with a 15 mM solution of tantalum (5) that had been converted to its PEG 550 (monomethyl ether) ester by exchange of ethyl alkoxides with PEG 550 in ethyl acetate;
  • subsequent treatment with 20 mM tetra(n-butyl)ammonium salt of tetraethylene glycol (monomethyl) monophosphate (TEG phosphate) in toluene at 110° C. for 4 days.
  • Element Atomic %
    Oxygen 54.4
    Carbon 25.3
    Tantalum 2.2
    Silicon 10.2
    Phosphorus 7.6
  • XPS data are shown in the table below for a titanium PEG-ester modified silicon dioxide surface after performing the following manipulations on the surface:
  • treatment of the quartz cover-slip for 18 hr. at methylene dichloride reflux with a 0.0002 M solution of titanium (4) that had been converted to its PEG 550 (monomethyl ether) ester by exchange of ethyl alkoxides with PEG 550 in methylene dichloride solvent;
  • subsequent treatment of the 0.005 M tetra(n-butyl)ammonium salt of tetraethylene glycol (monomethyl) monophosphate in methylene dichloride solvent at methylene dichloride reflux for 24 hr.
  • Element Atomic %
    Oxygen 53.8
    Carbon 24.4
    Titanium 2.3
    Silicon 15.8
    Phosphorus 3.5
  • The absorption of bio-molecules to these modified surfaces was tested by measuring the fluorescence from the surface after incubating with fluorescent labeled bio-molecules.
  • FIGS. 7A-7E are plots showing the results from confocal fluorescence imaging of coverslips with TaPEG550/TEG phosphate modified surfaces. The plots are histograms of the count rates in the pixels of a 100×100 μm image. FIG. 7A shows that, before any dye is added, the background fluorescence signal peaks around 2.5 kHz. FIGS. 7B and 7C show that when a 1 nM A488 solution is added, the count rate histogram peak shifts to 70 kHz and drops again back to signal levels comparable to the background when the dye is removed. This is repeated with a 1 nM R110 solution in the same 100×100 μm region as shown in FIG. 7D. Again, the results show that the fluorescence signals return to background levels when R110 is removed from the coverslip (FIG. 7E). These results demonstrate that the rhodamine dyes A488 and R110 do not non-specifically bind to the modified surface.
  • FIGS. 8A-8C are plots showing similar count rate histograms of a TaPEG550 (no phosphate) modified surface using sulfo-Liz dCTP which demonstrate that the modified surface prevents sulfo-Liz dCTP from nonspecifically binding to the coverslip. This is observed even when a high concentration sulfo-Liz dCTP solution is added onto the surface (FIG. 8B) and removed (FIG. 8C). Again, these results demonstrate that sulfo-Liz dCTP does not non-specifically bind to the modified surface.
  • FIGS. 9A-9C illustrate a comparison of the correlation curves obtained from fluorescence correlation spectroscopy (FCS) of dilute solutions of the above rhodamine dyes and the sulfoLiz dCTP in contact with TaPEG550 and silane PEG surface modified coverslips. The decay rate of the correlation curves provide information on the rate of diffusion of the free dyes in solution. Hence, if a dye sticks to the surface, the correlation curve will decay more slowly. For comparison, FIG. 9A is an example of a correlation curve exhibiting a tail at long lag times (e.g., 10 ms) indicative of sticky dyes at the surface. The data for FIG. 9A was obtained from 0.1 nM Liz-dGTP on conventionally prepared 5% PEGsilane modified coverslips wherein the PEGsilane was deposited from refluxing toluene.
  • FIG. 9B is a plot showing the correlation curves from dilute solutions of A488 and R110 diffusing above the modified surface. This plot also shows the correlation curve for freely diffusing A488 above a known non-sticky surface for comparison. FCS experiments were also done using dilute solutions of sulfo-Liz dCTP, the results of which are shown in FIG. 9C. This plot shows the results from three different FCS measurements at different positions on the modified coverslip. Together, these FCS results and the fluorescence imaging results above indicate that the TaPEG550 modified surfaces on fused silica substrate do not non-specifically bind A488, R110, and sulfo-Liz dCTP.
  • Treatment of Metal Reagent Modified Surfaces with Multi-Valent Passivating Agents
  • A quartz surface was treated with ethanolic Ta(monomethyl-PEG550)5 to form a modified surface. The modified surface was then treated with an ethanolic solution of a multivalent passivating agent having the following structure:
  • Figure US20110159305A1-20110630-C00011
  • which is designated penta(hydroxyl)-monomethyl-PEG550. The results are shown in FIGS. 10A-10D.
  • FIG. 10A shows non-specific binding data for 1 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550. FIG. 10B shows non-specific binding data for a 1 nm fluorescent LIZ dye solution treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550 after it had been incubated in a Tris buffer solution for 4 hours. FIG. 10C shows non-specific binding data for 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550. FIG. 10D shows non-specific binding data for a 100 nm fluorescent LIZ dye solution on a quartz surface treated with Ta(monomethyl-PEG550)5 followed by treatment with an ethanolic solution of penta(hydroxyl)monomethyl-PEG550 after the surface had been incubated in a Tris buffer solution for 4 hours. As can be seen from FIGS. 10A-10D, treatment with the multivalent passivating agent yields a surface that prevents non-specific adsorption of a 100 nM aqueous solution of Liz fluorescent dye even after 4 hours in the presence of a buffer.
  • In contrast, as shown in FIG. 11A-11C, some sticking of 1 nM of fluorescent Liz dye (FIG. 11A) and 100 nM of fluorescent Liz dye (FIG. 11B) was observed on Ta(monomethyl-PEG550)5 treated quartz slides which were not treated with the multivalent passivating agent. Extensive sticking was observed after 4 hours in the presence of a buffer (FIG. 11C).
  • The integrity of the Ta surface after treatment with the multivalent passivating agent (i.e., poly(hydroxyl)-monomethyl-PEG) is demonstrated by XPS of the modified quartz surfaces before and after poly(hydroxyl)-monomethyl-PEG modification, shown below.
  • XPS Data For Surface Treated With TaPEG550 Reagent
    Element Percentage
    Oxygen 63.19
    Carbon 9.43
    Tantalum 0.66
    Silicon 26.71
  • XPS Data For Surface Treated With TaPEG550 Reagent
    Followed By Poly(Hydroxyl)-Monomethyl-PEG
    Element Percentage
    Oxygen 52.3
    Nitrogen 2.5
    Carbon 26.8
    Tantalum 0.43
    Silicon 18.0
  • FIGS. 12A and 12B show fused silica surfaces treated with TaPEG550 then PEG350triphosphate before (FIG. 12A) and after (FIG. 12B) incubation in DNA extension buffer. The PEG350triphosphate has a chemical structure as set forth below:
  • Figure US20110159305A1-20110630-C00012
  • wherein n has a value such that the polyethylene glycol moiety has an average molecular weight of about 350. The treated surfaces were incubated with 100 nM LIZ-dATP and then rinsed with water to remove any dye that was not stuck to the surface. As can be seen from FIG. 12B, 100 nM LIZ-dATP exhibits minimal sticking to the modified surface even after the treated slide has been incubated with DNA extension buffer.
  • While the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be appreciated by one skilled in the art from reading this disclosure that various changes in form and detail can be made without departing from the true scope of the invention.

Claims (3)

1. A bio-molecule detection surface, comprising:
an oxide support material having a surface with hydroxyl and/or oxide anion groups, at least a portion of the hydroxyl and/or oxide anion groups bound to a metal reagent
wherein the metal reagent has a structure represented by the formula:

Y(L-Pol)m
wherein:
Y is a transition metal, magnesium or aluminum,
L is oxygen, sulfur, selenium or an amine,
each “Pol” group independently represents a polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, a substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and
m is an integer.
2. The bio-molecule detection surface of claim 1, wherein Y is zirconium.
3. The bio-molecule detection surface of claim 2, wherein L is oxygen, Pol is a hydrocarbon, and m is 1.
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Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7763423B2 (en) * 2005-09-30 2010-07-27 Pacific Biosciences Of California, Inc. Substrates having low density reactive groups for monitoring enzyme activity
US8975216B2 (en) * 2006-03-30 2015-03-10 Pacific Biosciences Of California Articles having localized molecules disposed thereon and methods of producing same
EP2069789A2 (en) * 2006-11-21 2009-06-17 Applied Biosystems, LLC Intermediates and methods for forming passivated surfaces on oxide layers and articles produced thereby
CN101459066B (en) * 2007-12-13 2010-08-11 中芯国际集成电路制造(上海)有限公司 Gate, shallow slot isolation region forming method and flattening method for silicon base etching surface
US8173198B2 (en) * 2008-07-23 2012-05-08 Life Technologies Corporation Deposition of metal oxides onto surfaces as an immobilization vehicle for carboxylated or phophated particles or polymers
US20100261185A1 (en) 2009-03-27 2010-10-14 Life Technologies Corporation Labeled enzyme compositions, methods and systems
US8501406B1 (en) 2009-07-14 2013-08-06 Pacific Biosciences Of California, Inc. Selectively functionalized arrays
WO2011106451A1 (en) * 2010-02-23 2011-09-01 Life Technologies Corporation Methods of treating metal oxide surfaces and surfaces produced thereby
US9366670B2 (en) 2011-07-04 2016-06-14 Samsung Electronics Co., Ltd. Polymer including group having at least two hydroxyls or zwitterionic group and use thereof
CN110431201A (en) * 2017-02-07 2019-11-08 昆塔波尔公司 Single polymer layer coating for modified metal-oxide surface
AU2018261007A1 (en) 2017-05-05 2019-11-07 Quantum-Si Incorporated Substrates having modified surface reactivity and antifouling properties in biological reactions
CN114829669A (en) 2019-10-11 2022-07-29 宽腾矽公司 Gas phase surface modification

Citations (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3770499A (en) * 1972-02-28 1973-11-06 Motorola Inc Liquid phase deposition of thin insulating and refractory film on a substrate
US6093370A (en) * 1998-06-11 2000-07-25 Hitachi, Ltd. Polynucleotide separation method and apparatus therefor
US6146767A (en) * 1996-10-17 2000-11-14 The Trustees Of Princeton University Self-assembled organic monolayers
US6277489B1 (en) * 1998-12-04 2001-08-21 The Regents Of The University Of California Support for high performance affinity chromatography and other uses
US20010029017A1 (en) * 1998-06-11 2001-10-11 Hitachi, Ltd. Polynucleotide separation method and apparatus therefor
US20020055146A1 (en) * 2000-03-10 2002-05-09 Shivashankar G. V. Control of the expression of anchored genes using micron scale heaters
US20020132242A1 (en) * 1997-04-16 2002-09-19 Gerdes John C. Nucleic acid archiving
US20020172963A1 (en) * 2001-01-10 2002-11-21 Kelley Shana O. DNA-bridged carbon nanotube arrays
US20030027328A1 (en) * 2000-10-30 2003-02-06 Sru Biosystems, Llc Guided mode resonant filter biosensor using a linear grating surface structure
US20030082658A1 (en) * 1998-04-10 2003-05-01 Francois Mallet Method for fixing a biological molecule on a support surface
US20030148401A1 (en) * 2001-11-09 2003-08-07 Anoop Agrawal High surface area substrates for microarrays and methods to make same
US6645644B1 (en) * 1996-10-17 2003-11-11 The Trustees Of Princeton University Enhanced bonding of phosphoric and phosphoric acids to oxidized substrates
US20040029162A1 (en) * 2000-12-20 2004-02-12 Patrick Chaton Support for determining an analyte comprising a selective spectral filter
US6811980B2 (en) * 2000-11-20 2004-11-02 Sony International (Europe) Gmbh Process for immobilization of nucleic acid molecules on a substrate
US20050003396A1 (en) * 2003-04-11 2005-01-06 Mihrimah Ozkan Biosensors having single reactant components immobilized over single electrodes and methods of making and using thereof
US6884628B2 (en) * 1999-04-28 2005-04-26 Eidgenossische Technische Hochschule Zurich Multifunctional polymeric surface coatings in analytic and sensor devices
US20050142296A1 (en) * 2003-12-30 2005-06-30 3M Innovative Properties Company Substrates and compounds bonded thereto
US20060035229A1 (en) * 2002-07-04 2006-02-16 Technische Universitat Dresden Metallic object with a nucleic acid coating and derivatives thereof and method for producing said object
US20060194008A1 (en) * 1999-09-22 2006-08-31 Princeton University Devices with multiple surface functionality
US20060205007A1 (en) * 2004-11-26 2006-09-14 Canon Kabushiki Kaisha DNA supporting fiber and DNA supporting fiber sheet and methods of producing them
US20060246225A1 (en) * 2003-05-20 2006-11-02 Thomas Moritz Sample carrier based on a porous film with metal oxide particles, the production and utilization thereof, especially for selective detection of phosphorylated/sulphatized biopolymers
US20070015223A1 (en) * 2003-11-28 2007-01-18 Olympus Corporation Method of detecting substance to be analyzed
US20070098892A1 (en) * 2005-08-31 2007-05-03 Samsung Electronics Co., Ltd. Method of forming a layer and method of manufacturing a capacitor using the same
US20070238679A1 (en) * 2006-03-30 2007-10-11 Pacific Biosciences Of California, Inc. Articles having localized molecules disposed thereon and methods of producing same
US20080020214A1 (en) * 2004-12-28 2008-01-24 Tomoji Kawai Method for immobilizing self-organizing material or fine particle on substrate, and substrate manufactured by using such method
US20080033867A1 (en) * 2002-12-09 2008-02-07 Creditex Group, Inc. Centralized process for determining deltas for index tranches
US20080044830A1 (en) * 2004-12-24 2008-02-21 Fraunhofer-Gesellschaft Zur Forderung Der Angewandten Forschung E.V. Three-Dimensional Nanostructured and Microstructured Supports
US20080132429A1 (en) * 2006-05-23 2008-06-05 Uchicago Argonne Biological microarrays with enhanced signal yield
US20080176761A1 (en) * 2006-11-21 2008-07-24 Applera Corporation, Applied Biosystems Group Intermediates and Methods for Forming Passivated Surfaces on Oxide Layers and Articles Produced Thereby
US20080274671A1 (en) * 2006-09-11 2008-11-06 Enbio Limited Method of doping surfaces
US7462452B2 (en) * 2004-04-30 2008-12-09 Pacific Biosciences Of California, Inc. Field-switch sequencing
US20090011949A1 (en) * 2001-07-11 2009-01-08 Hogan Michael E Methods and devices based upon a novel form of nucleic acid duplex on a surface
US20090156426A1 (en) * 2004-04-23 2009-06-18 Thomas Schiestel Functionalized porous supports for microarrays
US20090197777A1 (en) * 2007-08-13 2009-08-06 Alexander Chagovetz Method of making and using microarrays suitable for high throughput detection

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5789476A (en) 1995-03-03 1998-08-04 Seiko Epson Corporation Film-forming coating solution and synthetic resin lens
SE520046C2 (en) 1997-11-24 2003-05-13 Sense Ab Q Method for Measuring Interaction between a Target Molecule and a Receptor Molecule with Piezoelectric Crystal Microwave
GB9808264D0 (en) 1998-04-17 1998-06-17 Imperial College Biochemical devices and their methods of manufacture
DE60030978T2 (en) 1999-07-05 2007-06-14 Novartis Ag METHOD FOR USING A SENSOR UNIT
WO2001050131A1 (en) 2000-01-06 2001-07-12 Motorola, Inc. Three-dimensional network for biomolecule detection
AU2001289859A1 (en) 2000-09-05 2002-03-22 Zeptosens Ag Method for precipitating mono and multiple layers of organophosphoric and organophosphonic acids and the salts thereof in addition to use thereof
WO2003076903A2 (en) 2002-03-08 2003-09-18 Spectral Genomics, Inc. Articles of manufacture and methods for making hydrophobic derivatized arrays
WO2004011672A1 (en) 2002-07-26 2004-02-05 Ntera Limited Porous nanostructured film sensor
WO2004023128A1 (en) 2002-09-05 2004-03-18 Unisearch Limited Detection of target nucleic acid molecules by alteration of reaction of a redox species following hybridization with immoblized capture nucleic acid molecules
US7723126B2 (en) 2004-03-24 2010-05-25 Wisconsin Alumni Research Foundation Plasma-enhanced functionalization of inorganic oxide surfaces
EP1637613A1 (en) 2004-09-20 2006-03-22 Eidgenössische Technische Hochschule Zürich Electrochemical patterning on multi-channel microelectrode array for biosensing applications
JP2006177914A (en) * 2004-12-20 2006-07-06 Yukio Nagasaki Surface having thick polymer layer, and preparation method therefor
SE0403139D0 (en) 2004-12-23 2004-12-23 Nanoxis Ab Device and use thereof
EP1911844A1 (en) 2006-10-10 2008-04-16 Qiagen GmbH Methods and kit for isolating nucleic acids
CN101568835A (en) 2006-11-21 2009-10-28 应用生物系统公司 Intermediates and methods for forming passivated surfaces on oxide layers and articles produced thereby
WO2008063134A1 (en) 2006-11-24 2008-05-29 Agency For Science, Technology And Research Method of producing a pattern of discriminative wettability
JP2008266043A (en) 2007-04-17 2008-11-06 Tayca Corp Transparent titanium oxide sol and method for preparing the same
EP2011898B1 (en) 2007-07-03 2021-04-07 Beneq Oy Method in depositing metal oxide materials
WO2009129410A1 (en) 2008-04-18 2009-10-22 Cornell University Conformal particle coatings on fibrous materials

Patent Citations (46)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3770499A (en) * 1972-02-28 1973-11-06 Motorola Inc Liquid phase deposition of thin insulating and refractory film on a substrate
US6645644B1 (en) * 1996-10-17 2003-11-11 The Trustees Of Princeton University Enhanced bonding of phosphoric and phosphoric acids to oxidized substrates
US6146767A (en) * 1996-10-17 2000-11-14 The Trustees Of Princeton University Self-assembled organic monolayers
US20090082225A1 (en) * 1997-04-16 2009-03-26 Applera Corporation Nucleic acid archiving
US20020132242A1 (en) * 1997-04-16 2002-09-19 Gerdes John C. Nucleic acid archiving
US7087387B2 (en) * 1997-04-16 2006-08-08 Applera Corporation Nucleic acid archiving
US20040091925A1 (en) * 1997-04-16 2004-05-13 Gerdes John C. Nucleic acid archiving
US7361471B2 (en) * 1997-04-16 2008-04-22 Applera Corporation Nucleic acid archiving
US20030082658A1 (en) * 1998-04-10 2003-05-01 Francois Mallet Method for fixing a biological molecule on a support surface
US20010029017A1 (en) * 1998-06-11 2001-10-11 Hitachi, Ltd. Polynucleotide separation method and apparatus therefor
US20020048771A1 (en) * 1998-06-11 2002-04-25 Kenji Yasuda Polynucleotide separation meyhod and apparatus therefor
US20020055119A1 (en) * 1998-06-11 2002-05-09 Kenji Yasuda Polynucleotide separation method and apparatus therefor
US20020090641A1 (en) * 1998-06-11 2002-07-11 Hitachi, Ltd. Polynucleotide separation method and apparatus therefor
US6093370A (en) * 1998-06-11 2000-07-25 Hitachi, Ltd. Polynucleotide separation method and apparatus therefor
US6277489B1 (en) * 1998-12-04 2001-08-21 The Regents Of The University Of California Support for high performance affinity chromatography and other uses
US6884628B2 (en) * 1999-04-28 2005-04-26 Eidgenossische Technische Hochschule Zurich Multifunctional polymeric surface coatings in analytic and sensor devices
US20060194008A1 (en) * 1999-09-22 2006-08-31 Princeton University Devices with multiple surface functionality
US6489106B1 (en) * 2000-03-10 2002-12-03 Nec Research Institute, Inc. Control of the expression of anchored genes using micron scale heaters
US20020055146A1 (en) * 2000-03-10 2002-05-09 Shivashankar G. V. Control of the expression of anchored genes using micron scale heaters
US20030027328A1 (en) * 2000-10-30 2003-02-06 Sru Biosystems, Llc Guided mode resonant filter biosensor using a linear grating surface structure
US6811980B2 (en) * 2000-11-20 2004-11-02 Sony International (Europe) Gmbh Process for immobilization of nucleic acid molecules on a substrate
US20050266475A1 (en) * 2000-12-20 2005-12-01 Commissariat A L'energie Atomique Method for determining an analyte
US20040029162A1 (en) * 2000-12-20 2004-02-12 Patrick Chaton Support for determining an analyte comprising a selective spectral filter
US20020172963A1 (en) * 2001-01-10 2002-11-21 Kelley Shana O. DNA-bridged carbon nanotube arrays
US20090011949A1 (en) * 2001-07-11 2009-01-08 Hogan Michael E Methods and devices based upon a novel form of nucleic acid duplex on a surface
US20030148401A1 (en) * 2001-11-09 2003-08-07 Anoop Agrawal High surface area substrates for microarrays and methods to make same
US20060035229A1 (en) * 2002-07-04 2006-02-16 Technische Universitat Dresden Metallic object with a nucleic acid coating and derivatives thereof and method for producing said object
US7585965B2 (en) * 2002-07-04 2009-09-08 Technische Universitä{umlaut over ( )}t Dresden Metallic object with a nucleic acid coating and derivatives thereof and method for producing said object
US20080033867A1 (en) * 2002-12-09 2008-02-07 Creditex Group, Inc. Centralized process for determining deltas for index tranches
US20050003396A1 (en) * 2003-04-11 2005-01-06 Mihrimah Ozkan Biosensors having single reactant components immobilized over single electrodes and methods of making and using thereof
US20060246225A1 (en) * 2003-05-20 2006-11-02 Thomas Moritz Sample carrier based on a porous film with metal oxide particles, the production and utilization thereof, especially for selective detection of phosphorylated/sulphatized biopolymers
US20070015223A1 (en) * 2003-11-28 2007-01-18 Olympus Corporation Method of detecting substance to be analyzed
US20050142296A1 (en) * 2003-12-30 2005-06-30 3M Innovative Properties Company Substrates and compounds bonded thereto
US20090156426A1 (en) * 2004-04-23 2009-06-18 Thomas Schiestel Functionalized porous supports for microarrays
US7462452B2 (en) * 2004-04-30 2008-12-09 Pacific Biosciences Of California, Inc. Field-switch sequencing
US20060205007A1 (en) * 2004-11-26 2006-09-14 Canon Kabushiki Kaisha DNA supporting fiber and DNA supporting fiber sheet and methods of producing them
US20080044830A1 (en) * 2004-12-24 2008-02-21 Fraunhofer-Gesellschaft Zur Forderung Der Angewandten Forschung E.V. Three-Dimensional Nanostructured and Microstructured Supports
US20080020214A1 (en) * 2004-12-28 2008-01-24 Tomoji Kawai Method for immobilizing self-organizing material or fine particle on substrate, and substrate manufactured by using such method
US20070098892A1 (en) * 2005-08-31 2007-05-03 Samsung Electronics Co., Ltd. Method of forming a layer and method of manufacturing a capacitor using the same
US20080153100A1 (en) * 2006-03-30 2008-06-26 Pacific Biosciences Of California, Inc. Articles having localized molecules disposed thereon and methods of producing same
US20080032301A1 (en) * 2006-03-30 2008-02-07 Pacific Biosciences Of California, Inc. Articles having localized molecules disposed thereon and methods of producing same
US20070238679A1 (en) * 2006-03-30 2007-10-11 Pacific Biosciences Of California, Inc. Articles having localized molecules disposed thereon and methods of producing same
US20080132429A1 (en) * 2006-05-23 2008-06-05 Uchicago Argonne Biological microarrays with enhanced signal yield
US20080274671A1 (en) * 2006-09-11 2008-11-06 Enbio Limited Method of doping surfaces
US20080176761A1 (en) * 2006-11-21 2008-07-24 Applera Corporation, Applied Biosystems Group Intermediates and Methods for Forming Passivated Surfaces on Oxide Layers and Articles Produced Thereby
US20090197777A1 (en) * 2007-08-13 2009-08-06 Alexander Chagovetz Method of making and using microarrays suitable for high throughput detection

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