POLYMERIC SUBSTRATE WITH A SURFACE HAVING REDUCED BIOMOLECULE ADHESION, AND THERMOPLASTIC ARTICLES OF SUCH SUBSTRATE

Title:

POLYMERIC SUBSTRATE WITH A SURFACE HAVING REDUCED BIOMOLECULE ADHESION, AND THERMOPLASTIC ARTICLES OF SUCH SUBSTRATE

Document Type and Number:

WIPO Patent Application WO/2017/087032

Kind Code:

Abstract:

A substrate is described having a treated contact surface comprising a carbon or silicon compound comprising from 1 to 30 atomic percent oxygen, from 0.1 to 30 atomic percent nitrogen, or both, each as measured by XPS. The treated contact surface has a biomolecule recovery percentage greater than the biomolecule recovery percentage of the surface before treatment according to the method.

Inventors:

TAHA AHMAD (US)
FELTS JOHN T (US)

Application Number:

PCT/US2016/037674

Publication Date:

May 26, 2017

Filing Date:

June 15, 2016

Export Citation:

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Assignee:

SIO2 MEDICAL PRODUCTS INC (US)

International Classes:

C08J7/12; B01L3/00; C08J7/048; C08J7/056; C08J7/18

Domestic Patent References:

WO2013071138A1

2013-05-16

Foreign References:

EP2251452A2	2010-11-17
EP1859866A1	2007-11-28
US20040062882A1	2004-04-01
DE10066431B4	2013-10-24
US6068884A	2000-05-30
US4844986A	1989-07-04
US20060046006A1	2006-03-02
US20040267194A1	2004-12-30
US20140004022A1	2014-01-02
US20130209766A1	2013-08-15
US7985188B2	2011-07-26
US7399500B2	2008-07-15
US6828028B1	2004-12-07
US6828028B1	2004-12-07

Other References:

SPIERINGS ET AL: "Water absorption and the effects of moisture on the dynamic properties of synthetic mountaineering ropes", INTERNATIONAL JOURNAL OF IMPACT ENGINEERING, PERGAMON, GB, vol. 34, no. 2, 17 November 2006 (2006-11-17), pages 205 - 215, XP005771420, ISSN: 0734-743X, DOI: 10.1016/J.IJIMPENG.2005.08.008
J.V. DA MAIA ET AL: "Influence of gas and treatment time on the surface modification of EPDM rubber treated at afterglow microwave plasmas", APPLIED SURFACE SCIENCE, vol. 285, 1 November 2013 (2013-11-01), AMSTERDAM, NL, pages 918 - 926, XP055300824, ISSN: 0169-4332, DOI: 10.1016/j.apsusc.2013.09.013
MARK J. KUSHNER: "Pulsed Plasma- Pulsed Injection Sources For Remote Plasma Activated Chemical Vapor Deposition", J. A . P S., vol. 73, 1993, pages 4098, XP000885354, DOI: doi:10.1063/1.352840
S.J.LIMB; K.K.S.LAU; D.J.EDELL; E.F.GLEASON; K.K.GLEASON: "Molecular Design Of Fluorocarbon Film Architecture By Pulsed Plasma Enhanced And Pyrolytic Chemical Vapor Deposition", PLASMA POLYMERIZATION, vol. 4, 1999, pages 21 - 32, XP002541805, DOI: doi:10.1023/A:1021899414807
J. WANG; X. SONG; R. LIA; J. SHEN; G. YANG; H. HUANG: "Fluorocarbon Thin Film With Superhydrophobic Property Prepared By Pyrolysis Of Hexafluoropropylene Oxide", APPLIED SURFACE SCIENCE, vol. 258, 2012, pages 9782 - 9785

Attorney, Agent or Firm:

WHEELER, George F. (US)

Download PDF:

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Claims:

CLAIMS

1. A polymeric substrate consisting essentially of a contact surface and an interior portion, the contact surface comprising:

• from 1 to 30 atomic %, optionally from 5 to 20 atomic %, optionally from 5 to 10 atomic %, oxygen atoms,

• from 0 to 30 atomic %, optionally from 0 to 20 atomic %, optionally from 0 to 10 atomic %, nitrogen atoms, and

• from 70 to 99 atomic %, optionally from 80 to 95 atomic %, optionally from 90 to 95 atomic %, carbon atoms

in which the atomic % values are measured by x-ray photoelectron spectroscopy (XPS).

2. The substrate of any one preceding claim, comprising:

• from 1 to 30 atomic %, optionally from 5 to 20 atomic %, optionally from 5 to 10 atomic %, grafted oxygen atoms,

• from 0 to 30 atomic %, optionally from 0 to 20 atomic %, optionally from 0 to 10 atomic %, grafted nitrogen atoms.

3. The substrate of any one preceding claim, in which the interior portion comprises at least 80 atomic %, optionally at least 85 atomic %, optionally at least 90 atomic %, optionally at least 95 atomic %, optionally 100 atomic %, carbon atoms, and a larger atomic % of carbon atoms than the contact surface.

4. The substrate of any one preceding claim in which the contact surface comprises from 0.1 to 10 atomic %, optionally from 0.1 to 5 atomic %, optionally from 1 to 5 atomic %, carbon atoms to which oxygen is grafted.

5. The substrate of any one preceding claim in which the contact surface comprises from 0.1 to 10 atomic %, optionally from 0.1 to 5 atomic %, hydrogen bond acceptor groups.

6. The substrate of any one preceding claim, in which the grafted oxygen or nitrogen atoms are present in the form of moieties selected from -N⁺-, -N-, =0, -0-, -0₂, -0₃, C=0, 0-C=0, C-O-C, or

^≡

bonded to carbon or silicon in each case, optionally selected from or any combination of two or more of these.

7. The substrate of any one preceding claim, in which the contact surface comprises a carbon or silicon compound modified with polar grafted species, optionally one or more of: C=0 (carbonyl), C-O-C (ether), C-N-C (amine), C-N⁺-C (ammonium), C0₂ (carboxyl), C0₃, Si=0, Si-O-Si, Si-N-Si, Si-N⁺-Si, Si0₂, Si0₃,

or a combination of two or more of these, as measured by x-ray photoelectron spectroscopy (XPS).

8. The substrate of any one preceding claim, in which the contact surface comprises thermoplastic material, for example a thermoplastic resin, for example an injection-molded thermoplastic resin.

9. The substrate of any one preceding claim, in which the contact surface comprises a hydrocarbon polymer, for example an olefin polymer, polypropylene (PP), polyethylene (PE), cyclic olefin copolymer (COC), cyclic olefin polymer (COP), polymethylpentene, polystyrene, hydrogenated polystyrene, polycyclohexylethylene (PCHE), or combinations of two or more of these, or a heteroatom-substituted hydrocarbon polymer, for example a polyester, polyethylene terephthalate, polyethylene naphthalate, polybutylene terephthalate (PBT), polyvinylidene chloride (PVdC), polyvinyl chloride (PVC), polycarbonate, polylactic acid, epoxy resin, nylon, polyurethane polyacrylonitrile, polyacrylonitrile (PAN), an ionomeric resin, Surlyn® ionomeric resin, or any combination, composite or blend of any two or more of the above materials.

10. The substrate of any one preceding claim, in which the contact surface has a biomolecule recovery percentage of at least 10%, optionally at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 45%, optionally at least 50%, optionally at least 55%, optionally at least 60%, optionally at least 65%, optionally at least 70%, optionally at least 75%, optionally at least 80%, optionally at least 85%, optionally at least 90% optionally at least 95%.

11. The substrate of claim 10, in which the biomolecule recovery percentage is determined for at least one of: mammal serum albumin; Bovine Serum Albumin (BSA); Fibrinogen (FBG); Transferrin (TFN); egg white ovotransferrin (conalbumin); membrane-associated melanotransferrin; Protein A (PrA); Protein G (PrG); Protein A/G; Protein L; insulin; pharmaceutical protein; blood or blood component proteins; and any recombinant form, modification, full length precursor, signal peptide, propeptide, or mature variant of these proteins.

12. The substrate of claim 10, in which any one or more of the following is true:

• for BSA having an atomic mass of 66,000 Daltons, the biomolecule recovery percentage is at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 50%, optionally at least 60%, optionally at least 70%, optionally at least 80%, optionally at least 90%, optionally at least 95%, optionally at least 99%, optionally from 10% to 90%, optionally from 20% to 90%, optionally from 30% to 90%, optionally from 40% to 90%, optionally from 50% to 90%, optionally from 60% to 90%, optionally from 70% to 90%, optionally from 80% to 90%;

• for FBG having an atomic mass of 340,000 Daltons, the biomolecule recovery percentage is at least 10%, optionally at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 50%, optionally at least 60%, optionally at least 70%, optionally at least 80%, optionally at least 90%, optionally at least 95%, optionally at least 99%, optionally from 10% to 90%, optionally from 20% to 90%, optionally from 30% to 90%, optionally from 40% to 90%, optionally from 50% to 90%, optionally from 60% to 90%, optionally from 70% to 90%, optionally from 80% to 90%, optionally from 15% to 20%;

• for TFN having an atomic mass of 80,000 Daltons, the biomolecule recovery percentage is at least 30%, optionally at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 50%, optionally at least 60%, optionally at least 70%, optionally at least 80%, optionally at least 90%, optionally at least 95%, optionally at least 99%, optionally from 10% to 90%, optionally from 20% to 90%, optionally from 30% to 90%, optionally from 40% to 90%, optionally from 50% to 90%, optionally from 60% to 90%, optionally from 70% to 90%, optionally from 80% to 90%, optionally from 30% to 50%;

• for PrA having an atomic mass of 45,000 Daltons, the biomolecule recovery percentage is at least 10%, optionally at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 50%, optionally at least 60%, optionally at least 70%, optionally at least 80%, optionally at least 90%, optionally at least 95%, optionally at least 99%, optionally from 10% to 90%, optionally from 20% to 90%, optionally from 30% to 90%, optionally from 40% to 90%, optionally from 50% to 90%, optionally from 60% to 90%, optionally from 70% to 90%, optionally from 80% to 90%, optionally from 10% to 90%; and

• for PrG having an atomic mass of 20,000 Daltons, the biomolecule recovery percentage is at least 10%, optionally at least 20%, optionally at least 30%, optionally at least 40%, optionally at least 50%, optionally at least 60%, optionally at least 70%, optionally at least 80%, optionally at least 90%, optionally at least 95%, optionally at least 99%, optionally from 10% to 90%, optionally from 20% to 90%, optionally from 30% to 90%, optionally from 40% to 90%, optionally from 50% to 90%, optionally from 60% to 90%, optionally from 70% to 90%, optionally from 80% to 90%, optionally from 5% to 90%.

13. The substrate of any one preceding claim, in which the contact surface is a vessel lumen surface.

14. The substrate of any one preceding claim, in which the contact surface is a fluid contact surface of an article of labware.

15. The substrate of claim 14, in which the contact surface is a fluid contact surface of a microplate, a centrifuge tube, a pipette tip, a well plate, a microwell plate, an ELISA plate, a microtiter plate, a 96-well plate, a 384- well plate, a vial, a bottle, ajar, a syringe, a cartridge, a blister package, an ampoule, an evacuated blood collection tube, a specimen tube, a centrifuge tube, or a chromatography vial.

16. The substrate of any one preceding claim, in which the contact surface is in contact with an aqueous protein.

17. The substrate of any one preceding claim, in which the contact surface has a contact angle with distilled water of from 0° to 90°, optionally from 20° to 80°, optionally from 40° to 80°, optionally from 50° to 70°, optionally from 55° to 65°.

18. The substrate of any one preceding claim, in which the contact surface has an electrostatic charge build-up on the converted and optionally conditioned contact surface of less than 5 kilovolts per cm (5 kV/cm), optionally less than 3 kV/cm, optionally less than 2 kV/cm, optionally less than 1 kV/cm.

19. The substrate of any one preceding claim, in which the contact surface is treated with conditioning plasma, conversion plasma, or both of one or more non-polymerizing compounds to form the surface.

20. The substrate of Claim 19, in which the the non-polymerizing compound comprises, consists essentially of or consists of one or more of a member selected from the group consisting of: 0₂, N₂, air, 0₃, N₂0, H₂, H₂0, H₂0₂, NH₃, Ar, He and Ne.

21. The substrate of any one preceding claim, in which the contact surface is a PECVD deposited coating from 5 to 1000 nm thick.

22. The substrate of Claim 21, in which the PECVD deposited coating is applied using any one or combination of gas precursors including siloxane, organosiloxane, polyethylene glycol (PEG); or monomer gas precursors including acetylene, methane, propane, butane, ethane, and/or co-monomer gases including air, oxygen or water vapor.

23. The substrate of Claim 21, in which PECVD deposited coating is an amorphous carbon coating further comprising oxygen and optionally nitrogen heteroatoms.

24. The substrate of Claim 23, in which the amorphous carbon coating is applied using monomer gas precursors including acetylene, ethylene methane, ethane, propane, butane, or a combination of two or more of these and a co-monomer gas including air, oxygen, water vapor, or a combination of two or more of these.

25. A method of forming the polymeric substrate of any preceding claim, comprising • providing a polymeric article having a contact surface comprising carbon; and

• treating the contact surface with a conversion plasma comprising water vapor and oxygen under conditions effective to form a converted surface having a biomolecule recovery percentage, for an aqueous protein dispersion having a concentration from 0.01 nM to 1.4 nM, optionally 0.05 nM to 1.4 nM, optionally 0.1 nM to 1.4 nM, in contact with the converted surface, greater than the biomolecule recovery percentage of the contact surface before treating the contact surface with the conversion plasma.

26. The method of claim 25, in which the conditions of the treating step are effective to provide a biomolecule recovery percentage, for an aqueous protein dispersion having a concentration from 0.01 nM to 1.4 nM, optionally 0.05 nM to 1.4 nM, optionally 0.1 nM to 1.4 nM, in contact with the converted surface, greater than 80%.

27. The method of claim 25 or 26, in which the plasma is generated at a remote point from the contact surface, where the ratio of the radiant energy density at the remote point of conversion plasma treatment to the radiant energy density at the brightest point of the conversion plasma is less than 0.5, optionally less than 0.25, optionally substantially zero, optionally zero.

Description:

POLYMERIC SUBSTRATE WITH A SURFACE HAVING REDUCED BIOMOLECULE ADHESION, AND THERMOPLASTIC ARTICLES OF SUCH SUBSTRATE

[0001 ] This specification claims the priority of U. S . Provisional Patent Application Ser. No .

62/320,246, filed April 8, 2016. That application is incorporated here by reference in its entirety to provide continuity of disclosure.

[0002] The technology relates generally to a surface, or surface modification of a plastic substrate (sometimes referred to in this disclosure as a contact surface), having reduced biomolecule adhesion to the surface. More particularly, the technology relates to a plastic substrate, e.g., a medical device or item of laboratory ware, having reduced protein adhesion to the substrate surface.

[0003] In blood, biomolecule, and blood analyte testing, it is desirable to minimize biomolecule adsorption and binding to plastic ware used with these biological substances. Plastic microwell plates, chromatography vials, and other containers, as well as pipettes (sometimes spelled "pipets"), pipette tips, centrifuge tubes, microscope slides, and other types of laboratory ware (also known as labware) used to prepare and transfer samples commonly have hydrophobic surfaces and readily adsorb biomolecules such as proteins, DNA, and RNA. Surfaces of these and other types of laboratory ware components made of polymeric plastic can cause binding of the biomolecule samples. It is thus a desire to provide surfaces for plastic laboratory ware and other articles that contact biological substances, to reduce a wide range of biomolecules from adhering.

[0004] The present invention also relates to the technical field of fabrication of coated vessels for conducting chemical, biochemical, medical, and/or biological uses. These methods and systems are essential in a variety of applications including medical diagnostics, medical treatment, food and beverage testing, food and beverage packaging, environmental monitoring, manufacturing quality control, drug discovery, and scientific research. [0005] Laboratory in vitro and analytical testing is commonly carried out on special-purpose laboratory glassware and plastic ware to contain and locate the test samples. Traditionally glassware was used, and continues to be used, but glassware is readily breakable, very expensive, prone to particulate problems, yields heavy metal extractables, and can cause aggregation of proteins and other biologies.

[0006] Some of these problems can be addressed by substituting injection molded plastic ware for glassware. In particular, plastic ware is preferred in the biologies area, such as areas of medicine, medical research, drug discovery, and scientific research, due to the large number of issues with glassware. Plastic ware addresses some of the problems with glassware, but plastic ware creates certain problems as well. It can cause non-specific binding of biomolecules to the plastic ware as well as non-specific adsorption of biomolecules to the plastic ware. Plastic ware also contains leachables, preventing the use of plastic ware or making it undesirable for many types of Laboratory in vitro and analytical testing.

[0007] In addition, pipettes, pipette tips and centrifuge tubes are other types of laboratory ware used to prepare and transfer samples and transfer reagent solutions, and again can be glassware or plastic ware. In blood, biomolecule, and blood analyte testing, it is desirable to minimize biomolecule adsorption and non-specific biomolecule binding to the plastic ware. Further, it is desirable to minimize biomolecule adsorption and non-specific binding of biomolecules to the plastic ware in in vitro and analytical testing. Plastic microwell plates, vials, and other containers, as well as pipettes, pipette tips, centrifuge tubes, and other types of laboratory ware used to prepare and transfer samples having hydrophobic surfaces readily adsorb biomolecules such as proteins, DNA, and RNA. Surfaces of these and other types of laboratory ware components made of polymeric plastic can cause non-specific binding of the samples. Surfaces of these and other types of labware components made of glass can cause particulate problems, have heavy metal extractables, and can cause aggregation of proteins and other biomolecules. Previous hydrophilic coatings, including polyethylene glycol (PEG) and zwitterion polymeric coatings have been found to provide excellent biomolecule adsorption performance. Many of these polymeric coatings are not covalently bound to the article surface and have potential to move (dissolve, disperse) into the fluid payload causing interference with assay testing and/or exposing the plastic surface to protein surface adsorption, limiting their utility. Polymeric coating that are covalently attached to the article surface would not have the potential to move (dissolve, disperse) into the fluid pay load, eliminating this source of interference with assay testing. Further, covalently bound polymeric coating would prevent movement of the polymeric surface coating, thereby preventing undesired exposure of the article surface.

[0008] There is therefore a need for covalently bound polymeric coatings for the surface of plastic laboratory ware such as microwell plates, vials, other containers, pipettes, pipette tips, centrifuge tubes, and that will inhibit the adsorption of biomolecules to the surface of the plastic. Likewise, there is a need for bound polymeric coatings for the surface of plastic laboratory ware such as microwell plates, vials, other containers, pipettes, pipette tips, centrifuge tubes, and that will prevent movement of the polymeric surface coating thereby preventing undesired exposure of the plastic surface.

[0009] An important consideration in manufacturing packaging for regulated products, e.g., pharmaceuticals, is to ensure that the pharmaceutical product to be contained within a package is substantially free of contaminants. Therefore, processes for manufacturing and filling pharmaceutical packages with product, are typically performed under cleanroom conditions.

[0010] One cause of potential contamination is particulates. Particulate contamination may originate from various sources, which may be generally divided into two categories: (1) intrinsic contaminants; and (2) extrinsic contaminants. Intrinsic contaminants are product and process related particulates, for example, laser etching residues, filter media, cleanroom uniform fibers, rubber and plastic particles from filter housing, and needle shields. Extrinsic contamination comes from sources unrelated to product or process, for example, hair, skin cells, pollen, clothing fibers, salt and soil.

[0011] While filtration systems and good manufacturing practices can limit the surface and airborne particulate count in an area where containers are being manufactured or filled, these things do not always reduce particulate count on the container surfaces to acceptable levels. One particular challenge is presented by static charges of manufactured plastic containers, which tend to attract charged particles. Even if the airborne/surface particulate count is relatively low, a plastic container with a strong static charge can attract particulate contaminants and cause them to adhere to the container.

[0012] Attempted solutions to this problem include use of antistatic additives for polymers, such as ethoxylated alkylamines, ethoxylated alkyl amides, glycerol stearates, fatty acid esters, esters or ethers of polyols and sodium alkyl sulfonates. The amounts of such additives in polymers typically vary from 0.1 % to 3 % by weight. While these additives are somewhat effective in reducing the static charges of plastic articles that incorporate them, the additives are mobile in the polymer matrix and tend to bloom to the surface. Additives that bloom to the surface can contaminate the surface and the contents, especially liquid contents, of a container made from a polymer with such additives.

[0013] There is therefore a need for plastic articles that are treated to reduce their static charge without the use of typical antistatic additives, which may themselves be a source of contamination. Likewise, there is a need for methods of treating plastic articles to reduce their static charge without the use of typical antistatic additives. Additionally, many of these coatings require fluid phase chemistry, comprising reactants and solvents.

SUMMARY

[0014] An aspect of the invention is a polymeric substrate consisting essentially of a contact surface and an interior portion. The contact surface includes from 1 to 30 atomic oxygen atoms, from 0 to 30 atomic % nitrogen atoms, and from 70 to 99 atomic % carbon atoms. The atomic % values are measured by x-ray photoelectron spectroscopy (XPS).

[0015] Another aspect of the invention is a method of forming the substrate of any preceding claim by providing a polymeric article having a contact surface comprising carbon and treating the contact surface. The contact surface is treated with a conversion plasma comprising water vapor and oxygen. The treatment is carried out under conditions effective to form a converted surface having a biomolecule recovery percentage greater than the biomolecule recovery percentage of the contact surface before treating the contact surface with the conversion plasma. The biomolecule recovery percentage is determined for an aqueous protein dispersion having a concentration from 0.01 nM to 1.4 nM, optionally 0.05 nM to 1.4 nM, optionally 0.1 nM to 1.4 nM, in contact with the converted surface.

[0016] Other aspects of the invention overcome the deficiencies of prior coatings and coated vessels by providing covalently bound polymeric zwitterionic monolayers or epoxy-modified zwitterion-containing copolymeric monolayers which can provide excellent inhibition of surface absorption of biomolecules and inhibition of non-specific binding of biomolecules with the surface without concern for polymer dissolution/dispersion into the fluid media. Embodiment (i) provides the reaction of a surface-bound hydroxyl group with a siloxane-functional polymer containing zwitterion functionality. Embodiment (ii) provides the reaction of a hydroxylated plastic or SiOx- coated glass surface with an epoxy-modified zwitterion-containing copolymers to assure covalent surface bonding.

[0017] The invention utilizes covalent bond formation between a hydroxylated plastic or

SiOx-coated glass surface and, in Embodiment (i) a zwitterion- siloxane copolymer, resulting from condensation (water or volatile alcohol loss (e.g., surface SiOH + polymer (zwitterion-siloxane- Si(OR)x)→ surface SiO-Si (polymer (zwitterion-siloxane))) + ROH or in Embodiment (ii) an epoxy-modified zwitterion-containing copolymer, resulting from functionalization of a hydroxylated plastic or SiOx-coated glass surface with a silyl amine coupling agent followed by further functionalization with a zwitterionic epoxy copolymer to form a epoxy-modified zwitterion- containing copolymeric monolayer.

[0018] The invention generally relates to a method for coating a surface of a vessel, such as laboratory ware and vessels used, e.g., for purposes of analytical chemistry, medical research, drug discovery, and scientific research (e.g., chromatography vials, microplates, pipettes, pipette tips, centrifuge test tubes, and the like), or other vessels and/or surfaces that come into contact with biomolecule containing substances such as beakers, round bottom flasks, Erlenmeyer flasks, graduated cylinders, and the like. Biomolecules may include, for example, simple biomolecules (e.g., nucleotides, amino acids, simple sugars and fatty acids), nucleic acid molecules, polynucleotides, polypeptides, polysaccharides, lipids, steroids, or a conjugate of these (e.g., a glycoprotein, a lipoprotein, a glycolipid, DNA-protein conjugate), antibodies, receptors, receptor fusion constructs, receptor fusion proteins, chimeric proteins, antibody fusion constructs, antibody fusion proteins, as well as cells, viruses, and bacteria. The preferred application of the present invention is with, for example, non-parenteral containers, although parenteral containers treated according to processes disclosed herein are also within the scope.

[0019] More particularly, the present invention relates to, for example, covalently bound (i) polymeric zwitterionic monolayers or (ii) epoxy-modified zwitterion-containing copolymeric monolayers, which have been known to show superior resistance to non-specific biomolecule, such as protein, adsorption and/or non-specific binding in the laboratory environment, where the zwitterionic deposition has been done using multiple step processes. Here, a commercially viable process offering both low cost and product performance has been developed based on the concept of the unique biomolecule, such as protein, adsorption resistance feature of covalently bound (i) polymeric zwitterionic monolayer coatings or (ii) epoxy-modified zwitterion-containing copolymeric monolayer. Covalently bound (i) polymeric zwitterionic monolayer coatings or (ii) epoxy-modified zwitterion-containing copolymeric monolayer coatings for these products improve their sample- contact characteristics, for example to adapt them for contact with biomolecule containing samples, such as protein or other peptide or polypeptide samples, to provide a surface which has, for example, non-binding or low-binding properties. For example, the present invention contemplates methods for coating surfaces of these products with a covalently bound (i) polymeric zwitterionic monolayer coatings or (ii) epoxy-modified zwitterion-containing copolymeric monolayer coating, to yield a nonspecific biomolecule adsorption and/or non-specific binding resistant surface coating. These covalently bound (i) polymeric zwitterionic monolayer coatings or (ii) epoxy-modified zwitterion- containing copolymeric monolayer coatings have favorable properties when used as a coating for surfaces that come into contact with biomolecule containing samples.

[0020] Additionally, in one aspect, the invention is a method of reducing static charge of a plastic container. The invention utilizes the reaction of a (i) surface-bound hydroxyl group or (ii) hydroxylated plastic or SiOx-coated glass surface with a siloxane-functional polymer containing zwitterion functionality to provide covalently bound (i) polymeric zwitterionic monolayer coatings or (ii) epoxy-modified zwitterion-containing copolymeric monolayers which can provide excellent biomolecule adsorption inhibition and non-specific biomolecule surface binding inhibition without concern for polymer dissolution/dispersion into the fluid media. The coating reduces static charge of the container compared to a reference container that is essentially identical to the container except that the reference container is uncoated.

[0021] The invention utilizes sequential reactions.

[0022] In Embodiment (i), these sequential reactions involve (1) utilizing a hydroxy- modified or an SiOx plasma-modified surface followed by (2) forming a zwitterion-siloxane copolymer, resulting from condensation (water or volatile alcohol loss (e.g. surface SiOH + polymer (zwitterion-siloxane-Si(OR)x)→ surface SiO-Si (polymer (zwitterion-siloxane))) + ROH. In certain embodiments, a hydroxyl-functional surface can be (A) formed, e.g., from 02/plasma reaction onto a carbon-based polymer surface (AC), forming C-OH moieties, or onto a silicon-based (Si02 or silicone) surface (ASi), forming Si-OH moieties. Additionally, hydroxyl-functional surfaces can be (B) inherent to the nascent surface, e.g., polymers such as polyvinyl alcohol (PVOH) or silicon-based polymer surfaces (Si02 or silicones). Thus the 02/plasma surface preparation method can be optional if inherent hydroxyl moieties are present on the surface.

[0023 ] In Embodiment (ii) these sequential reactions involve ( 1 ) forming a hydroxy-modified or an SiOx plasma-modified surface followed by (2) forming an amine modified surface by reacting the hydroxy-modified or an SiOx plasma-modified surface with a silyl amine coupling agent followed by (3) forming an amine linked by condensation of the amine modified surface with an epoxy-modified zwitterion-containing copolymer.

[0024] The coating may be both polymeric and hydrophilic (binds water to ionic moieties) providing a durable steric and bound-water barrier to biomolecule adsorption and non-specific biomolecule binding onto plastic surfaces.

[0025] While the zwitterionic layers may be thinner than preformed polymeric zwitterionic coatings, solvent coating methods, thru evaporation, can cause non-uniform coatings, resulting in (a) optically observable nonuniformities, (b) wasted coating (more coating thickness than required/desired), and the inventive materials should provide good biomolecule adsorption and/or binding inhibition.

[0026] A non-exhaustive list of patents of possible relevance includes U.S. Pat. Nos.

6,068,884 and 4,844,986 and U.S. Published Applications 20060046006, 20040267194, 20140004022, and 20130209766.

[0027] All references cited herein are incorporated herein by reference in their entireties.

[0028] The invention provides a method for providing a non-specific biomolecule adsorption and/or binding resistant coated surface which exhibits low- or non-binding of biomolecules and/or low or non-adsorption of biomolecules.

[0029] Embodiment (i) of the method comprises the steps of: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02 in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a hydroxyl functional substrate surface, and optionally heating the substrate; (e) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises at least one siloxane moiety and at least one zwitterion moiety; (f) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (g) optionally waiting; (h) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or non-specific binding resistant coated surface.

[0030] Embodiment (ii) of the method comprises the steps of (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02, optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; (h) optionally waiting; (i) removing the copolymer, and optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or non-specific binding resistant coated surface.

[0031] Either embodiment (i) or (ii) provides a method for providing a non-specific biomolecule adsorption and/or non-specific binding resistant coated surface which exhibits low- or non-binding of biomolecules and/or low or non-adsorption of biomolecules.

[0032] Embodiment (i) comprises the steps of: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising an organosilicon precursor and optionally 02 in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a thus forming a hydroxyl functional substrate surface; (e) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises at least one siloxane moiety and at least one zwitterion moiety; (f) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (g) optionally waiting; (h) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated surface.

[0033] Embodiment (ii) comprises the steps of: (a) providing a substrate that is a SiOx- coated glass surface reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02, optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; (h) removing the copolymer solution, (i) optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, and (j) optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated surface.

[0034] Embodiment (i) provides a method for providing a non-specific biomolecule adsorption and/or non-specific binding resistant coated surface which exhibits low- or non-binding of biomolecules and/or low or non-adsorption of biomolecules, the method comprising the steps: (a) providing a substrate with a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (d) optionally waiting;(e) removing the copolymer solution, and optionally drying the nonspecific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated surface.

[0035] Embodiment (i) provides a method wherein the zwitterionic copolymer binds covalently to the prepared substrate surface.

[0036] Embodiment (ii) provides a method wherein the at least one silyl amine coupling agent is selected from the group consisting of 3-Aminopropyltriethoxysilane, 3- Aminopropyltrimethoxysilane, 4-Aminobutyltriethoxysilane, M-Aminophenyltrimethoxysilane, P- Aminophenyltrimethoxysilane, O-Aminophenyltrimethoxysilane, 3-

Aminopropyltris(Methoxyethoxyethoxy)Silane, 11-Aminoundecyltriethoxysilane, 3-(M- Aminophenoxy)Propyltrimethoxysilane, Aminopropylsilanetriol, 3-

Aminopropylmethyldiethoxysilane, 3 - Aminopropyldiisopropylethoxysilane, 3 -

Aminopropyldimethylethoxysilane, N-(2- Aminoethyl)-3 - Aminopropyltrimethoxy silane, N- [3 - (Trimethoxysilyl)Propyl]Ethylenediamine, N-(2-Aminoethyl)-3-Aminopropyltriethoxysilane, N-(6-Aminohexyl)Aminomethyltriethoxysilane, N-(6-Aminohexyl)Aminopropyltrimethoxysilane, N-(2-Aminoethyl)- 11-Aminoundecyltrimethoxysilane,

Phenylaminopropyltrimethoxysilane, 3-(N-Allylamino)Propyltrimethoxysilane, (Cyclohexylaminomethyl)Triethoxysilane, N-Cyclohexylaminopropyltrimethoxysilane, N- Ethylaminoisobutylmethyldiethoxysilane, (Phenylaminomethyl)Methyldimethoxysilane, N- Phenylaminomethyltriethoxysilane, N-Methylaminopropylmethyldimethoxysilane, Bis(2- Hydroxyethyl)-3-Aminopropyltriethoxysilane, 3-(N-Styrylmethyl-2-Aminoethylamino)- Propyltrimethoxysilane Hydrochloride, (2-N-Benzylaminoethyl)-3 - Aminopropyl-Trimethoxysilane, Bis(Triethoxysilylpropyl)Amine, Bis(Trimethoxysilylpropyl)Amine, Bis[(3-

Trimethoxy silyl)Propyl] - Ethylenediamine, Bis [(3 -Trimethoxysilyl)Propyl] -Ethylenediamine, Bis(Methyldiethoxysilylpropyl)Amine, N-Allyl-Aza-2,2-Dimethoxysila- Cyclopentane, N- Aminoethyl-Aza-2,2,4-Trimethyl-Silacyclopentane, N-(3-Aminopropyldimethylsila)Aza-2,2- Dimethyl-2-Silacyclopentane, N-N-Butyl-Aza-2,2-Dimethoxysila- Cyclopentane, 2,2-Dimethoxy- l,6-Diaza-2-Silacyclo-Octane, N-Methyl-Aza-2,2,4-Trimethylsila-Cylcopentane, 1-(N-(N',N'- dimethylaminoethyl))-l-aza-2,2,4-trimethyl-2-silacyclopentan e and combinations thereof.

[0037] Embodiment (ii) provides a method wherein the at least one copolymerizable zwitterionic monomer is selected from the group consisting of 2-(meth)acryloyloxyethyl-2'- (trimethylammonio)ethyl phosphate (including MPC), 2-(methacryloxy)ethyl 2- (diimethylammonio)ethyl phosphate (aka = 2-methacryloyloxyethyl phosphorylcholine; 2- (methacryloxy)ethyl-2-(dimethylammonio)ethylcarboxylate; 2-(methacryloxy)ethyl-2- (dimethylammonio)propylsulfonate; 2-(methyacrylamido)ethyl-2- (dimethylammonio)propylsulfonate; phosphoric acid 2-(methacryloyloxy)ethyl 2- (trimethylammonio) ethyl ester); 3-(meth)acryloyloxypropyl-2'-(trimethylammonio)ethyl phosphate, 4-(meth)acryloyloxybutyl-2'-(trimethylammonio)ethyl phosphate 5-(meth)acryloyloxypentyl-2'- (trimethylammonio)ethyl phosphate, 6-(meth)acryloyloxyhexyl-2'-(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-(trimethylammonio)ethyl phosphate, 2-

(meth)acryloyloxyethyl-2'-(tripropylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-

(tributylammonio)ethyl phosphate, 2-(meth)acryloyloxypropyl-2'-(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxybutyl-2'-(trimethylammonio)ethyl phosphate, 2-

(meth)acryloyloxypentyl-2'-(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyhexyl-2'-

(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-3'-(trimethylammonio)propyl phosphate, 3-(meth)acryloyloxypropyl-3'-(trimethylammonio)propyl phosphate, 4-

(meth)acryloyloxybutyl-3'-(trimethylammonio)propyl phosphate, 5-(meth)acryloyloxypentyl-3'-

(trimethylammonio)propyl phosphate, 6-(meth)acryloyloxyhexyl-3'-(trimethylammonio)propyl phosphate, 2-(meth)acryloyloxyethyl-4'-(trimethylammonio)butyl phosphate, 3-

(meth)acryloyloxypropyl-4'-(trimethylammonio)butyl phosphate, 4-(meth)acryloyloxybutyl-4'-

(trimethylammonio)butyl phosphate, 5-(meth)acryloyloxypentyl-4'-(trimethylammonio)butyl phosphate, 6-(meth)acryloyloxyhexyl-4'-(trimethylammonio)butyl phosphate, 2-(vinyloxy)ethyl-2'-

(trimethylammonio)ethyl phosphate, 2-(allyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(p- vinylbenzyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(p-vinylbenzoyloxy)ethyl-2'-

(trimethylammonio)ethyl phosphate, 2-(styryloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(p- vinylbenzyl)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(vinyloxycarbonyl)ethyl-2'-

(trimethylammonio)ethyl phosphate, 2-(allyloxycarbonyl)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(acryloylamino)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-

(vinylcarbonylamino)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(allyloxycarbonylamino)ethyl-

2'-(trimethylammonio)ethyl phosphate, 2-(butyroyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate,

2-(crotonoyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, ethyl-(2'- trimethylammonioethylphosphorylethyl) fumarate, butyl - (2'- trimethylammonioethylphosphorylethyl) fumarate, hydroxyethyl - (2'- trimethylammonioethylphosphorylethyl)fumarate, 2-hydroxyethyl(meth)acrylate, 2- hydroxypropyl(meth)acrylate; polyalkylene glycol(meth)acrylate monomers diethylene glycol mono(meth)acrylate, polyethylene glycol mono(meth)acrylate, polypropylene glycol mono(meth)acrylate, methoxydiethylene glycol(meth)acrylate, methoxypolyethylene glycol(meth)acrylate, methoxypolypropylene glycol(meth)acrylate; ro-carboxy(meth)acrylates, phthalic acid monohydroxyethyl(meth)acrylate, hexahydrophthalic acid monohydroxyethyl(meth)acrylate, ω-carboxy-polycaprolactone (n=2 to 5) mono(meth)acrylate, (meth)acryloyloxyethyl succinate; N-vinyl-2-pyrrolidone, vinylpyridine, (meth)acrylamide, (meth)acrylic acid, 2-acrylamide-2-methylpropanesulfonic acid, dimethylaminoethyl(meth)acrylate, diethylaminoethyl(meth)acrylate, maleic anhydride, and glyco-2-hydroxyethyl monomethacrylate (GEMA), D-carboxyethyl-3,3-dimethyammoniumethylmethacylate and sulfopropyl-3,3- dimethyammoniumethylmethacylate, and combinations thereof.

[0038] Embodiment (ii) provides a method wherein the at least one copolymerizable epoxy monomer is selected from the group consisting of glycidyl acrylate, glycidyl methacrylate (GMA), (E)-(oxiran-2-yl)methyl but-2-enoate, (3,3-dimethyloxiran-2-yl)methyl methacrylate, (E)-(oxiran-2- yl)methyl cinnamate, (oxiran-2-yl)methyl 2-methylenebutanoate, l-(oxiran-2-yl)propyl acrylate, 1- (oxiran-2-yl)ethyl methacrylate, (oxiran-2-yl)methyl 3-methyl-2-methylenebutanoate, (oxiran-2- yl)methyl 2-methylenepentanoate, (3-methyloxiran-2-yl)methyl acrylate, 2-(oxiran-2-yl)propan-2-yl acrylate, (oxiran-2-yl)methyl 2-methylenehexanoate, (3 -methyloxiran-2-yl)methyl methacrylate, and (3,3-dimethyloxiran-2-yl)methyl acrylate and combinations thereof.

[0039] Embodiment (ii) provides a method wherein the epoxy-modified zwitterionic copolymer binds covalently to the amine modified substrate.

[0040] In either embodiment (i) or (ii), the solvent is selected from the group consisting of water, methanol, ethanol, isopropanol, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, N,N- dimethylformamide, dimethylsulfoxide, methylene chloride, 1,2-dicholorethane and combinations thereof.

[0041 ] Either embodiment (i) or (ii) provides a method wherein the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface has decreased binding of biomolecules compared to a non-treated substrate.

[0042] Either embodiment (i) or (ii) provides a method wherein the biomolecule is selected from the group consisting of nucleotides, amino acids, sugars, fatty acids, nucleic acid molecules, polypeptides, polynucleotides, polysaccharides, lipids, steroids, glycoproteins, lipoproteins, glycolipids, DNA, DNA-protein conjugates, RNA, RNA-conjugates, antibodies, and receptors, receptor fusion constructs, receptor fusion proteins, chimeric proteins, antibody fusion constructs, and antibody fusion proteins.

[0043] Either embodiment (i) or (ii) provides a method wherein the substrate is a surface of a vessel.

[0044] Either embodiment (i) or (ii) provides a method wherein the vessel is selected from the group consisting of a microplate; a centrifuge tube; a pipette tip; a cuvette, a microwell plate; an ELISA plate; a microtiter plate; a 96-well plate; a 384-well plate; a 1536-well plate; a round bottom flask; and an Erlenmeyer flask.

[0045] Embodiment (i) provides, as one alternative, a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate made by: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising an organosilicon precursor and optionally 02 in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a hydroxyl functional substrate surface; (e) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises at least one siloxane moiety and at least one zwitterion moiety; (f) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (g), optionally waiting; (h) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated substrate.

[0046] Embodiment (i) further provides, as another alternative, a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate made by: (a) providing a substrate with a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (d) optionally waiting; (e) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated substrate.

[0047] Embodiment (ii) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate made by: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02 optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated substrate.

[0048] Embodiment (i) provides a non-specific biomolecule adsorption and/or binding resistant coated substrate wherein the non-specific biomolecule adsorption and/or binding resistant coating has decreased binding of biomolecules compared to a non-treated substrate. The biomolecule is selected from the group consisting of nucleotides, amino acids, sugars, fatty acids, nucleic acid molecules, polypeptides, polynucleotides, polysaccharides, lipids, steroids, glycoproteins, lipoproteins, glycolipids, DNA, DNA-protein conjugates, RNA, RNA-conjugates, antibodies, and receptors, receptor fusion constructs, receptor fusion proteins, chimeric proteins, antibody fusion constructs, and antibody fusion proteins.

[0049] Embodiment (ii) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate made by: (a) providing a substrate that is a SiOx-coated glass surface reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02, optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; (h) optionally waiting; (i) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated substrate.

[0050] In Embodiment (ii), the at least one silyl amine coupling agent is selected from the group consisting of 3-Aminopropyltriethoxysilane, 3-Aminopropyltrimethoxysilane, 4- Aminobutyltriethoxysilane, M-Aminophenyltrimethoxysilane, P-Aminophenyltrimethoxysilane, O- Aminophenyltrimethoxysilane, 3-Aminopropyltris(Methoxyethoxyethoxy)Silane, 11- Aminoundecyltriethoxysilane, 3-(M-Aminophenoxy)Propyltrimethoxysilane, Aminopropylsilanetriol, 3-Aminopropylmethyldiethoxysilane, 3-

Aminopropyldiisopropylethoxysilane, 3-Aminopropyldimethylethoxysilane, N-(2-Aminoethyl)-3- Aminopropyltrimethoxysilane, N- [3 -(Trimethoxysilyl)Propyl] Ethylenediamine, N-(2-Aminoethyl)- 3-Aminopropyltriethoxysilane, N-(6-Aminohexyl)Aminomethyltriethoxysilane, N-(6-

Aminohexyl) Aminopropyltrimethoxysilane, N-(2- Aminoethyl)- 11 - Aminoundecyltrimethoxysilane, (Aminoethylaminomethyl)Phenethyltrimethoxysilane, N-3-[(Amino(Polypropylenoxy)], Aminopropyltrimethoxysilane, N-(2-Aminoethyl)-3-Aminopropylsilanetriol, N-(2-Aminoethyl)-3- Aminopropylmethyldimethoxysilane, N-(2-Aminoethyl)-3-Aminoisobutylmethyldimethoxysilane, (Aminoethylamino)-3-Isobutyldimethylmethoxysilane, (3- Trimethoxysilylpropyl)Diethylenetriamine, N-Butylaminopropyltrimethoxysilane, N- Ethylaminoisobutyltrimethoxysilane, N-Methylaminopropyltrimethoxysilane, N-

[0051 ] Embodiment (ii) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate wherein the at least one copolymerizable zwitterionic monomer is selected from the group consisting of 2-(meth)acryloyloxyethyl-2'-(trimethylammonio)ethyl phosphate (including MPC), 2-(methacryloxy)ethyl 2-(diimethylammonio)ethyl phosphate (aka = 2- methacryloyloxyethyl phosphorylcholine; 2-(methacryloxy)ethyl-2-

(dimethylammonio)ethylcarboxylate; 2-(methacryloxy)ethyl-2-(dimethylammonio)propylsulfonate; 2-(methyacrylamido)ethyl-2-(dimethylammonio)propylsulfonate; phosphoric acid 2- (methacryloyloxy)ethyl 2-(trimethylammonio) ethyl ester); 3-(meth)acryloyloxypropyl-2'- (trimethylammonio)ethyl phosphate, 4-(meth)acryloyloxybutyl-2'-(trimethylammonio)ethyl phosphate 5-(meth)acryloyloxypentyl-2'-(trimethylammonio)ethyl phosphate, 6- (meth)acryloyloxyhexyl-2'-(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'- (trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-(tripropylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-(tributylammonio)ethyl phosphate, 2-

(meth)acryloyloxyhexyl-3'-(trimethylammonio)propyl phosphate, 2-(meth)acryloyloxyethyl-4'-

(trimethylammonio)butyl phosphate, 3-(meth)acryloyloxypropyl-4'-(trimethylammonio)butyl phosphate, 4-(meth)acryloyloxybutyl-4'-(trimethylammonio)butyl phosphate, 5-

(meth)acryloyloxypentyl-4'-(trimethylammonio)butyl phosphate, 6-(meth)acryloyloxyhexyl-4'-

(trimethylammonio)butyl phosphate, 2-(vinyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-

(allyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(p-vinylbenzyloxy)ethyl-2'-

(trimethylammonio)ethyl phosphate, 2-(p-vinylbenzoyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(styryloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(p-vinylbenzyl)ethyl-2'-

(trimethylammonio)ethyl phosphate, 2-(vinyloxycarbonyl)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(allyloxycarbonyl)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(acryloylamino)ethyl-

2'-(trimethylammonio)ethyl phosphate, 2-(vinylcarbonylamino)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(allyloxycarbonylamino)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-

(butyroyloxy)ethyl-2'-(trimethylammonio)ethyl phosphate, 2-(crotonoyloxy)ethyl-2'-

(meth)acryloyloxyethyl succinate; N-vinyl-2-pyrrolidone, vinylpyridine, (meth)acrylamide,

(meth)acrylic acid, 2-acrylamide-2-methylpropanesulfonic acid, dimethylaminoethyl(meth)acrylate, diethylaminoethyl(meth)acrylate, maleic anhydride, and glyco-2-hydroxyethyl monomethacrylate

(GEMA), b-carboxyethyl-3,3-dimethyammoniumethylmethacylate and sulfopropyl-3,3- dimethyammoniumethylmethacylate, and combinations thereof. [0052] Embodiment (i) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate made by: (a) providing a substrate with a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (d), optionally waiting; (e) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated substrate.

[0053] Embodiment (i) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate wherein the zwitterionic copolymer binds covalently to the prepared substrate surface

[0054] The at least one copolymerizable epoxy monomer of Embodiment (ii) is selected from the group consisting of glycidyl acrylate, glycidyl methacrylate (GMA), (E)-(oxiran-2-yl)methyl but-2-enoate, (3,3-dimethyloxiran-2-yl)methyl methacrylate, (E)-(oxiran-2-yl)methyl cinnamate, (oxiran-2-yl)methyl 2-methylenebutanoate, l-(oxiran-2-yl)propyl acrylate, l-(oxiran-2-yl)ethyl methacrylate, (oxiran-2-yl)methyl 3-methyl-2-methylenebutanoate, (oxiran-2-yl)methyl 2- methylenepentanoate, (3-methyloxiran-2-yl)methyl acrylate, 2-(oxiran-2-yl)propan-2-yl acrylate, (oxiran-2-yl)methyl 2-methylenehexanoate, (3-methyloxiran-2-yl)methyl methacrylate, and (3,3- dimethyloxiran-2-yl)methyl acrylate and combinations thereof.

[0055] Either embodiment (i) or (ii) provides a non-specific biomolecule adsorption and/or binding resistant coated substrate wherein the non-specific biomolecule adsorption and/or binding resistant coating has decreased binding of biomolecules compared to a non-treated substrate.

[0056] Either embodiment (i) or (ii) provides a non-specific biomolecule adsorption and/or binding resistant coated substrate wherein the biomolecule is selected from the group consisting of nucleotides, amino acids, sugars, fatty acids, nucleic acid molecules, polypeptides, polynucleotides, polysaccharides, lipids, steroids, glycoproteins, lipoproteins, glycolipids, DNA, DNA-protein conjugates, RNA, RNA-conjugates, antibodies, and receptors, receptor fusion constructs, receptor fusion proteins, chimeric proteins, antibody fusion constructs, and antibody fusion proteins.

[0057] The invention provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate wherein the substrate is a surface of a vessel.

[0058] The invention provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated substrate wherein the vessels is selected from the group consisting of a microplate; a centrifuge tube; a pipette tip; a cuvette, a microwell plate; an ELIS A plate; a microtiter plate; a 96-well plate; and a 384-well plate.

[0059] Embodiment (i) in one alternative provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02 in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a hydroxyl functional substrate surface, and optionally heating the substrate; (e) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (f) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (g) optionally waiting; (h) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0060] Embodiment (i) in another alternative provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (c) optionally waiting; (d) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0061] Embodiment (ii) provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02, optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; (h) optionally waiting; (i) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0062] Embodiment (i) provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate in a reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising an organosilicon precursor and optionally 02 in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a hydroxyl functional surface on the substrate surface; (e) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (f) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (g) optionally waiting; (h) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel. [0063] Embodiment (ii) provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate in a that is a SiOx-coated glass surface reaction chamber; (b) drawing a vacuum in the reaction chamber; (c) providing a gas comprising 02, optionally containing argon, optionally containing nitrogen in the vicinity of the substrate surface; (d) generating a plasma from the gas, thus forming a prepared substrate surface, and optionally heating the substrate; (e) providing a vapor comprising at least one silyl amine coupling agent to provide an amine modified substrate; (f) providing a copolymer solution comprising a solvent, wherein the copolymer comprises at least one copolymerizable zwitterionic monomer and at least one copolymerizable epoxy monomer, optionally in the presence of a polymerization initiator; (g) reacting the copolymer with the amine modified substrate and optionally heating the substrate; (h) optionally waiting; (i) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or non-specific binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0064] Embodiment (i) provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (c) optionally waiting; (d) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0065] Embodiment (i) alternatively provides a vessel having an interior surface coated at least in part with a non-specific biomolecule adsorption and/or binding resistant coating made by: (a) providing a substrate a hydroxyl functional surface; (b) providing a copolymer solution comprising a solvent and at least one zwitterionic copolymer, wherein the at least one zwitterionic copolymer comprises two or more monomers, comprising at least one siloxane moiety and at least one zwitterion moiety; (c) reacting the hydroxyl functional surface with the at least one zwitterionic copolymer and optionally heating the substrate; (c) optionally waiting; (d) removing the copolymer solution, and optionally drying the non-specific biomolecule adsorption and/or binding resistant coated surface, optionally heating the surface, thus forming a non-specific biomolecule adsorption and/or binding resistant coated vessel.

[0066] The invention provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated vessel wherein the zwitterionic copolymer binds covalently to the prepared substrate surface.

[0067] Embodiment (ii) provides a non-specific biomolecule adsorption and/or binding resistant coated vessel wherein the at least one silyl amine coupling agent is selected from the group consisting of 3-Aminopropyltriethoxysilane, 3-Aminopropyltrimethoxysilane, 4- Aminobutyltriethoxysilane, M-Aminophenyltrimethoxysilane, P-Aminophenyltrimethoxysilane, O- Aminophenyltrimethoxysilane, 3-Aminopropyltris(Methoxyethoxyethoxy)Silane, 11- Aminoundecyltriethoxysilane, 3-(M-Aminophenoxy)Propyltrimethoxysilane, Aminopropylsilanetriol, 3-Aminopropylmethyldiethoxysilane, 3-

Aminopropyldiisopropylethoxysilane, 3-Aminopropyldimethylethoxysilane, N-(2-Aminoethyl)-3- Aminopropyltrimethoxysilane, N- [3 -(Trimethoxysilyl) Propyl] Ethylenediamine, N-(2-Aminoethyl)- 3-Aminopropyltriethoxysilane, N-(6-Aminohexyl)Aminomethyltriethoxysilane, N-(6-

Aminohexyl) Aminopropyltrimethoxysilane, N-(2- Aminoethyl)- 11 - Aminoundecyltrimethoxysilane, (Aminoethylaminomethyl)Phenethyltrimethoxysilane, N-3-[(Amino(Polypropylenoxy)], Aminopropyltrimethoxysilane, N-(2-Aminoethyl)-3-Aminopropylsilanetriol, N-(2-Aminoethyl)-3- Aminopropylmethyldimethoxysilane, N-(2-Aminoethyl)-3-Aminoisobutylmethyldimethoxysilane, (Aminoethylamino)-3-Isobutyldimethylmethoxysilane, (3-Trimethoxysilylpropyl) Diethylenetriamine, N-Butylaminopropyltrimethoxysilane, N-Ethylaminoisobutyltrimethoxysilane, N-Methylaminopropyltrimethoxysilane, N-Phenylaminopropyltrimethoxysilane, 3-(N- Allylamino)Propyltrimethoxysilane, (Cyclohexylaminomethyl)Triethoxysilane, N-

Cyclohexylaminopropyltrimethoxysilane, N-Ethylaminoisobutylmethyldiethoxysilane, (Phenylaminomethyl)Methyldimethoxysilane, N-Phenylaminomethyltriethoxysilane, N-

Methylaminopropylmethyldimethoxysilane, Bis(2-Hydroxyethyl)-3-Aminopropyltriethoxysilane, 3- (N-Styrylmethyl-2-Aminoethylamino)- Propyltrimethoxysilane Hydrochloride, (2-N- Benzylaminoethyl)-3-Aminopropyl-Trimethoxysilane, Bis(Triethoxysilylpropyl)Amine, Bis(Trimethoxysilylpropyl)Amine, Bis[(3-Trimethoxysilyl)Propyl]- Ethylenediamine, Bis [(3- Trimethoxysilyl) Propyl] - Ethylenediamine, Bis(Methyldiethoxysilylpropyl)Amine, N-Allyl-Aza- 2,2-Dimethoxysila- Cyclopentane, N-Aminoethyl-Aza-2,2,4-Trimethyl-Silacyclopentane, N-(3- Aminopropyldimethylsila)Aza-2,2-Dimethyl-2-Silacyclopentane, N-N-Butyl-Aza-2,2- Dimethoxysila- Cyclopentane, 2,2-Dimethoxy-l,6-Diaza-2-Silacyclo-Octane, N-Methyl-Aza-2,2,4- Trimethylsila-Cylcopentane, l-(N-(N',N'-dimethylaminoethyl))-l-aza-2,2,4-trimethyl-2- silacyclopentane and combinations thereof.

[0068] Embodiment (ii) provides a non-specific biomolecule adsorption and/or non-specific binding resistant coated vessel wherein the at least one copolymerizable zwitterionic monomer is selected from the group consisting of 2-(meth)acryloyloxyethyl-2'-(trimethylammonio)ethyl phosphate (including MPC), 2-(methacryloxy)ethyl 2-(diimethylammonio)ethyl phosphate (aka = 2- methacryloyloxyethyl phosphorylcholine; 2-(methacryloxy)ethyl-2-(dimethylammonio) ethylcarboxylate; 2-(methacryloxy)ethyl-2-(dimethylammonio)propylsulfonate; 2- (methyacrylamido)ethyl-2-(dimethylammonio)propylsulfonate; phosphoric acid 2- (methacryloyloxy)ethyl 2-(trimethylammonio) ethyl ester); 3-(meth)acryloyloxypropyl-2'- (trimethylammonio)ethyl phosphate, 4-(meth)acryloyloxybutyl-2'-(trimethylammonio)ethyl phosphate 5-(meth)acryloyloxypentyl-2'-(trimethylammonio)ethyl phosphate, 6- (meth)acryloyloxyhexyl-2'-(trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'- (trimethylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-(tripropylammonio)ethyl phosphate, 2-(meth)acryloyloxyethyl-2'-(tributylammonio)ethyl phosphate, 2-