JP2023533742A - Modification of metal surfaces with phosphonic acid - Google Patents

Abstract

Provided herein are methods and reagents for the functionalization of metal surfaces, including platinum, gold, palladium, iridium, and rhodium. The methods and reagents provided herein use phosphonic acid or phosphonate ester reagents as attachment sources for various functional groups, which can modify surfaces with desired properties. The modified surfaces provided herein are useful in a variety of applications, including preventing biofouling on metal surfaces. [Selection drawing] Fig. 1A

Description

Cross-Reference This application claims the benefit of US Provisional Application No. 63/050,228, filed July 10, 2020, which is hereby incorporated by reference in its entirety.

Surface modification of the material can be used to achieve different interfacial properties of the material. For example, an antimicrobial layer, a corrosion resistant layer, or an anti-biofouling layer can be incorporated into the surface of a medical device designed to be implanted within the body. Although many chemistries exist for the covalent modification of various surfaces, few methods are known for the functionalization of platinum, gold, or other metal surfaces such as palladium, iridium, or rhodium. . A need exists for additional approaches for attaching reagents and other functional groups to such metal surfaces.

Provided herein are reagents, methods, and compositions that meet the need for new and improved methods of functionalizing metal surfaces, including platinum and gold surfaces. In some embodiments, the metal surface is a platinum group metal. In some embodiments, the reagents, methods, and compositions can be used to functionalize palladium, iridium, or rhodium surfaces. In some embodiments described herein, the reagents, methods, and compositions are used to attach various compounds to metal surfaces through the formation of metal-phosphonate bonds between phosphonic acid or phosphonate esters and metal surfaces. Functional groups can be added. In some embodiments, the phosphonic acid or phosphonate is attached to the metal surface with additional substances, such as additional surface modifying groups (e.g., hydrogels) or other functional moieties (e.g., binding agents such as peptides or oligonucleotides). contain reactive groups or trapping substances that allow the addition of In other embodiments, the phosphonic acid or phosphonate includes desired surface-modifying groups (eg, without having to perform a separate reaction to add the desired groups). In some embodiments, the metal surface is a platinum surface or its oxide. In some embodiments, the metal surface is a gold surface or its oxide. In some embodiments, the metal surface is a palladium surface or its oxide. In some embodiments, the metal surface is an iridium surface or its oxide. In some embodiments, the metal surface is a rhodium surface or its oxide.

The reagents, methods, and compositions of the present disclosure are used to enable precise control of attachment of various functional groups to metal surfaces, including platinum, gold, palladium, iridium, rhodium, or their oxides. particularly suitable. The materials provided herein allow for facile functionalization of surfaces with controlled amounts of reagents that form stable bonds with metal surfaces. The ease of synthesis, the robustness of the synthetic protocol, and the stability of the metal-phosphonate bonds allow for easy and controlled further functionalization of the surface. In some embodiments, this allows for the development of robust protocols for adding additional surface functionalization groups in a manner that allows fine control of properties. For example, hydrogels with tunable properties can be polymerized on metal surfaces functionalized with the reagents provided herein. Fine tuning of hydrogel properties such as density, thickness, crosslink density, graft density, functional groups, or conductivity can be performed as the reagents and protocols provided herein provide predictable results. A similar process can be used to develop robust protocols for functionalization with other surface modification groups.

Also provided herein are hydrogel materials suitable for grafting onto metal surfaces to prevent surface biofouling. Such hydrogels are particularly useful when deployed in electrodes and other electrokinetic devices that generate an electric field in close proximity to a biological sample. This is because the existence of the hydrogel layer can prevent sample destruction and accumulation of deteriorated sample on the electrode surface. In order for such a hydrogel layer to perform this function without interfering with the performance of current flow on the sample, the hydrogel layer is constructed with certain superior properties (e.g., density, thickness, and conductivity). is important, which requires precise control over the formation of the hydrogel layer on the surface. Therefore, there is a need for reagents that can functionalize surfaces and allow controlled deposition of surface-modifying materials, including hydrogels. In addition to being applied to platinum, gold, palladium, iridium, or rhodium surfaces as described herein, these hydrogels can also be used on silicon, aluminum, titanium, iron, zinc, zirconium, nickel, silver, copper Other metal surfaces capable of forming bonds with phosphonates can also be linked, including but not limited to, cobalt, or chromium.

In one aspect, provided herein is a structure
A functionalized metal surface comprising a metal surface bound to a molecule having
During the ceremony,
R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted and the metal surface is selected from platinum, gold, palladium, iridium, and rhodium, oxides thereof, or any combination thereof. Contains one metal.

In some embodiments, R has the structure XL- and L is a linking moiety and X is a reactive handle or capture moiety, or R has the structure YL- and L is the linking moiety and Y is the surface modifying group. In some embodiments, L is an optionally substituted alkylene chain or an optionally substituted heteroalkylene chain.
In some embodiments, L is of the formula -[Z-(CR ¹ R ² ] _n ] _m -
has the structure of
wherein each R ¹ and R ² is independently H or optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted or optionally substituted heteroaryl, or R ¹ and R ² on the same atom taken together form oxo, or R ¹ on adjacent atoms taken together forming a double bond or R ¹ and R ² on adjacent atoms together form a triple bond,
Each Z does not exist independently, -O-, -NR ³ -, -S-, -SS-, -S(O)-, -S(O) ₂ -, -C(O)-, - C(O)O—, —C(O)NR ³ , —OC(O)O—, —NR ³ C(O)NR ³ —, —OC(O)NR ³ —, —S(O) ₂ O -, -S(O) ₂ NR ³ -, -OS(O) ₂ O-, -NR ³ SO ₂ NR ³ -, or -OSO ₂ NR ³ -;
Each R ³ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl,
Each n is independently an integer from 1-30, and m is an integer from 1-30.

In some embodiments, L comprises a chain of 1-100 atoms, each atom in said chain being independently selected from C, N, O, S, Si, and P. In some embodiments, the first atom in the chain is C.

In some embodiments, L is
wherein each n is independently an integer from 1-30. In some embodiments, L is the structure
wherein each n is independently an integer from 1-30 and each p is independently an integer from 1-30.

In some embodiments, L comprises a polymer. In some embodiments, the polymer is polyethylene, polypropylene, poly(vinyl halide), polystyrene, nylon, polyamide, polyester, polyaramid, polyacrylate, polymethacrylate, polytetrafluoroethylene, polysaccharide, poly(alkylene oxide ), poly(vinylpyrrolidone), poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine), or any combination or derivative thereof.

In some embodiments, R has the structure XL-. In some embodiments, X is azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, acid, activated ester, alkene, alcohol, halide, acyl halide, sulfonic acid, sulfinic acid, sulfonyl halides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, and hydrazides, or any combination thereof. In some embodiments, X has the structure X ¹ -L ^X -Z ^X -
has
In the formula, Z ^X is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)-, -C( O)O—, —C(O)NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O—, —S(O) ₂ NR ⁴ —, —OS(O) ₂ O—, —NR ⁴ S(O) ₂ NR ⁴ —, or —OS(O) ₂ NR ⁴ —;
L ^X is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted arylene , optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene;
X ¹ is an azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, carboxylic acid, active ester, alkene, alcohol, acyl halide, sulfonic acid, sulfinic acid, sulfonyl halide, epoxide, aldehyde, ketone , imines, oximes, isocyanates, isothiocyanates, hydrazines, or hydrazides, and each ^R4 is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl , optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

In some embodiments, Z ^X is a bond, -O-, -C(O)-, -NR ⁴ -, -C(O)O-, or -C(O)NR ⁴ . In some embodiments, L ^X has the structure —(CR ⁵ R ⁶ ) _r —, wherein each R ⁵ and R ⁶ is independently H, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl, or ^R5 and ^R6 on the same atom together form oxo, or adjacent atoms ^R5 together form a double bond, or adjacent atoms ^R5 and ^R6 together form a triple bond , and r is an integer from 0-30. In some embodiments, X ¹ comprises an amine, carboxylic acid, maleimide, azide, alkyne, halide, alkene, or sulfhydryl.

In some embodiments, X is
and
During the ceremony,
R ⁵ and R ⁶ are each independently H or C ₁ -C ₃ alkyl;
r is 0 or 1 and X ¹ is an azide, alkyne, halide, alkene, or sulfhydryl.

In some embodiments, X is
is. In some embodiments, X comprises a capture moiety selected from biotin, avidin, streptavidin, nucleic acids, peptides, and proteins, or any combination thereof.

In one aspect, provided herein is a kit comprising a functionalized metal surface provided herein and instructions for use.

In some embodiments, R has the structure YL-. In some embodiments, Y is a hydrophobic residue, hydrophilic residue, charged residue, cationic residue, anionic residue, polysaccharide, hydrophobic polymer, hydrophilic polymer, antimicrobial agent, biomaterial , biocompatible materials, antifouling materials, conductive materials, semiconducting materials, heat resistant materials, anticorrosion materials, catalysts, and magnetic materials, or any combination thereof.

In some embodiments, Y has the structure Y ¹ -L ^Y -Z ^Y -,
In the formula, Z ^Y is a bond, —O—, —NR ₄ —, —S—, —SS—, —S(O)—, S(O) ₂ —, —C(O)—, —C(O ) O—, —C(O)NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O—, — S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -, or azide, alkyne, tetrazine, halide, sulfhydryl , disulfides, maleimides, amines, carboxylic acids, activated esters, alkenes, alcohols, acyl halides, sulfonic acids, sulfinic acids, sulfonyl halides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, or a reaction product formed by a covalent bond-forming reaction between a hydrazide and a suitable complementary reactive group;
L ^Y is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted arylene, optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene;
Y ¹ contains at least one surface modification residue, and each R ⁴ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

In some embodiments, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)- , —C(O)O—, —C(O)NR ⁴ —, —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O ) ₂ O-, -S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -,
is. In some embodiments, ^LY is absent. In some embodiments Y ¹ comprises a hydrophobic residue. In some embodiments, the hydrophobic residues comprise hydrophobic polymers. In some embodiments, the hydrophobic polymer is polyethylene, polypropylene, polystyrene, polyvinyl halide, polytetrafluoroethylene, polymethylmethacrylate, polycarbonate, polyether-urethane, polydimethylsiloxane, or any combination or derivative thereof. including. In some embodiments the hydrophobic residue comprises a fatty acid or derivative thereof.

In some embodiments Y ¹ comprises a hydrophilic residue. In some embodiments, the hydrophilic residue comprises a hydrophilic polymer. In some embodiments, the hydrophilic polymer is polyacrylamide, polyacrylate, polymethacrylate, poly(alkylene oxide), poly(vinylpyrrolidone), poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine) , or any combination of derivatives thereof. In some embodiments, the hydrophilic polymer is polyacrylamide, poly(diethylacrylamide), poly(dimethylacrylamide), poly(N-isopropylacrylamide), poly(acrylic acid), poly(methacrylic acid), poly( methyl acrylate), poly(ethyl acrylate), poly(2-hydroxyethyl acrylate), poly(propyl acrylate), poly(butyl acrylate), poly(methyl methacrylate), poly(2-hydroxyethyl methacrylate), poly(tetrahydrofuran furyl methacrylate), poly(ethylene oxide), poly(propylene oxide), poly(vinylpyrrolidone), polyoxazoline, poly(2-ethyloxazoline), or poly(N-acryloylmorpholine).

In some embodiments, Y is the structure
including, where
each R ⁷ is independently H, optionally substituted C ₁ -C ₃ alkyl, or optionally substituted C ₁ -C ₃ hydroxyalkyl;
each R ⁸ is independently H or C ₁ -C ₃ alkyl;
each X ⁷ is independently absent, -O-, -S-, or -NR ⁹ ;
Each R ⁹ is independently H or C ₁ -C ₆ alkyl, and s is an integer from 1-10,000.

In some embodiments, Y ¹ comprises a cationic residue. In some embodiments, Y ¹ comprises a cationic polymer. In some embodiments, the cationic residues comprise protonated amine groups, protonated substituted amine groups, quaternary amine groups, or any combination thereof. In some embodiments, Y ¹ comprises an anionic residue. In some embodiments, Y ¹ comprises an anionic polymer. In some embodiments, the anionic residues comprise carboxylates, sulfonates, sulfinates, phosphates, phosphonates, or any combination thereof.

In some embodiments, the surface-modifying residues are hydrophobic residues, hydrophilic residues, charged residues, cationic residues, anionic residues, polysaccharides, hydrophobic polymers, hydrophilic polymers, antimicrobial residues. agents, biomaterials, biocompatible materials, antifouling materials, conductive materials, semiconducting materials, heat resistant materials, anti-corrosion materials, catalysts, magnetic materials, or any combination thereof. In some embodiments, the surface-modifying group comprises hydrogel. In some embodiments, the hydrogel comprises acrylate, methacrylate, acrylamide, or methacrylamide, or any combination thereof. In some embodiments, the hydrogel has a thickness of about 0.001 microns to about 10 microns. In some embodiments, the hydrogel has a conductivity of about 0.1 S/m to about 10 S/m.

In some embodiments, at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50% of the metal surface , at least about 60%, at least about 70%, at least about 80%, or at least about 90% functionalized. In some embodiments, the metal surface is at least partially oxidized. In some embodiments, at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50% of the metal surface , is at least about 60%, at least about 70%, at least about 80%, or at least about 90% oxidized.

In some embodiments, the metal surface comprises a metal alloy. In some embodiments, the metal alloy comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% by weight of the metal. In some embodiments, the metal surface comprises metal and additional materials. In some embodiments, the metal surface is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% by weight of a metal or any oxidation of a metal. Including things.

In some embodiments, the metal surface is configured to be placed inside a mammal. In some embodiments, the metal surface is a surgical or dental implant. In some embodiments, the metal surface is an electrode, microchip, bead, microparticle, or nanoparticle.

In some embodiments, the metal is platinum or gold, or oxides thereof. In some embodiments, the metal is palladium, iridium, or rhodium, or oxides thereof.

In one aspect, provided herein is a method of functionalizing a metal surface, the method comprising the steps of (a) depositing a phosphonic acid or phosphonate ester reagent on the metal surface; heating the metal surface to bind a phosphonic acid or phosphonate ester reagent to the metal surface, wherein the metal surface is platinum, gold, palladium, iridium, or rhodium, oxides thereof; or any combination thereof.

In some embodiments, depositing the phosphonic acid or phosphonate ester reagent comprises contacting the metal surface with a solution comprising the phosphonic acid reagent and a solvent. In some embodiments, the solvent comprises organic solvent, aqueous solvent, or any combination or mixture thereof. In some embodiments, the organic solvent is acetic acid, acetone, acetonitrile, benzene, tert-butyl alcohol, tert-butyl methyl ether, carbon tetrachloride, chloroform, cyclohexane, 1,2-dichloroethane, dichloromethane, diethyl ether, Diglyme, 1,2-dimethoxyethane, dimethylacetamide, dimethylformamide, dimethylsulfoxide, dioxane, ethanol, ethyl acetate, ethyl methyl ketone, ethylene glycol, hexanes, hexamethylphosphoramide, methanol, nitromethane, pentanes, 2-proponal , pyridine, tetrahydrofuran, toluene, xylene, or any combination thereof. In some embodiments, the organic solvent comprises ethanol, tetrahydrofuran, or toluene.

In some embodiments, the phosphonate or phosphonate ester reagent is present in solution at concentrations up to about 1 μM, 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1 M. In some embodiments, attaching the phosphonic acid or phosphonate ester reagent further comprises evaporating the solvent from the metal surface. In some embodiments, evaporating the solvent from the metal surface comprises heating the metal surface. In some embodiments, the metal surface is heated to a temperature of at least 30°C, at least 40°C, at least 50°C, at least 60°C, at least 70°C, at least 80°C, or at least 90°C. In some embodiments, heating the metal surface to bind the phosphonic acid reagent to the metal surface is performed in an oven, vacuum oven, or microwave reactor. In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface heats the metal surface to at least 30° C., at least 50° C., at least 70° C., at least 80° C. °C, at least 100 °C, at least 120 °C, at least 140 °C, at least 160 °C, or at least 180 °C.

In some embodiments, the metal surface is a metal oxide surface. In some embodiments, the metal oxide surface is oxidized by air oxidation, plasma treatment, ultraviolet ozone oxidation, or chemical oxidation. Some embodiments further comprise oxidizing the metal surface. In some embodiments, oxidizing the metal surface comprises plasma treatment, ultraviolet ozone oxidation, or chemical oxidation.

In some embodiments, the phosphonic acid or phosphonate ester reagent has the structure
and where
R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl substituted with or optionally substituted heteroaryl;
Each R ^E is independently H or substituted alkyl.

In some embodiments, the metal surface comprises platinum or gold, or oxides thereof. In some embodiments, the metal surface comprises palladium, iridium, or rhodium, or oxides thereof.

In one aspect, provided herein is a metal surface microarray comprising probe moieties that bind to the metal surface via phosphonate residues, wherein the metal surface comprises platinum, gold, palladium, iridium, or rhodium, or oxides thereof, or any combination thereof. In some embodiments, the attached phosphonate residue has the structure
where R comprises a probe moiety and each

indicates the point of attachment to the surface.

In some embodiments, the probe moieties comprise nucleic acids, peptides, proteins, antibodies, small molecules, glycans, or any combination thereof. In some embodiments, the probe portion comprises nucleic acids. In some embodiments, the nucleic acid comprises DNA or RNA. In some embodiments, microarrays are configured for DNA or RNA sequencing. In some embodiments, the probe moieties bind to biological agents. In some embodiments, the biological agent is a nucleic acid, protein, cell, or organelle. In some embodiments, the probe moieties are specific to biological agents. In some embodiments, the microarray comprises at least 10, at least 100, at least 1000, at least 10000, or at least 100000 unique probe features. In some embodiments, the metal surface microarray comprises platinum or gold, or oxides thereof. In some embodiments, the metal surface microarray comprises palladium, iridium or rhodium, oxides thereof, or any combination thereof.

In one aspect, provided herein is a drug delivery device comprising a drug moiety linked to a surface of the device via a phosphonate residue, wherein the surface is platinum, gold, palladium, iridium, or A drug delivery device comprising rhodium, oxides thereof, or combinations thereof. In some embodiments, the phosphonate residue has the structure

wherein R comprises a drug moiety and each

indicates the point of attachment to the surface.

In some embodiments, the drug moiety is linked to the phosphonate residue via a linker. In some embodiments, the linker is a cleavable linker. In some embodiments, the linker is configured to release drug moieties. In some embodiments, the drug delivery device further comprises a targeting moiety. In some embodiments, the targeting moiety comprises an antibody or antibody fragment, peptide, or nucleic acid. In some embodiments, the drug delivery device is a nanoparticle, microparticle, or bead. In some embodiments, the surface comprises platinum or gold, oxides thereof, or combinations thereof. In some embodiments, the surface comprises palladium, iridium, or rhodium, oxides thereof, or combinations thereof.

In one aspect, provided herein is a structure

A functionalized metal surface comprising a metal surface that binds to a molecule having
where R is the metal surface, including the hydrogel.

In some embodiments, the metal surface is platinum, gold, palladium, iridium, rhodium, silicon, aluminum, titanium, iron, zinc, zirconium, nickel, silver, copper, cobalt, or chromium, or oxides thereof. , or any combination thereof.

In some embodiments, the hydrogel has a conductivity of about 0.001 S/m to about 10 S/m. In some embodiments, the hydrogel has a thickness of about 0.001 microns to about 10 microns. In some embodiments, the hydrogel comprises synthetic polymers. In some embodiments, the hydrogel comprises an acrylamide polymer. In some embodiments, the acrylamide polymer comprises N-substituted acrylamide, N-substituted methacrylamide, or methacrylamide, or any combination thereof. In some embodiments, the hydrogel is polyacrylamide, poly(diethylacrylamide), poly(dimethylacrylamide), poly(N-isopropylacrylamide), poly(acrylic acid), poly(methacrylic acid), poly(methyl acrylate). ), poly(ethyl acrylate), poly(2-hydroxyethyl acrylate), poly(propyl acrylate), poly(butyl acrylate), poly(methyl methacrylate), poly(2-hydroxyethyl methacrylate), poly(tetrahydrofurfuryl methacrylate ), poly(ethylene oxide), poly(propylene oxide), poly(vinylpyrrolidone), polyoxazoline, poly(2-ethyloxazoline), or poly(N-acryloylmorpholine). In some embodiments, the metal surface is an electrode. In some embodiments, the metal surface is disposed on the outer surface of the electrode.

INCORPORATION BY REFERENCE All publications, patents, and patent applications mentioned in this specification are identified as if the individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. is incorporated herein by reference to the same extent.

The novel features of the invention are set out with particularity in the appended claims. A better understanding of the features and advantages of the present invention may be had by reference to the following detailed description and accompanying drawings, which set forth illustrative embodiments in which the principles of the invention are employed.
An example reaction of a phosphonic acid reagent with a metal surface is shown. Two exemplary bonding modes of phosphonate modified metal surfaces are shown. An example workflow for modifying a metal surface is shown. X-ray photoelectron spectroscopy measurements of oxidized and untreated platinum tips are shown. Figure 2 shows the oxidation state of oxidized and untreated platinum chips measured by X-ray photoelectron spectroscopy. Optical profilometer measurements of unfunctionalized bare platinum electrodes are shown. Optical profilometer measurements of platinum electrodes functionalized with phosphonic acid reagents are shown. Figure 2 shows a scanning electron microscope (SEM) image of a platinum electrode surrounded by dielectric material at a tilt angle of 45°. SEM images of a functionalized platinum electrode and subsequent polymerization reaction are shown. SEM image of a polymer-functionalized platinum electrode at a tilt angle of 0°. SEM images of polymer-functionalized platinum electrodes at a tilt angle of 14° are shown. SEM images of polymer-functionalized platinum electrodes at a tilt angle of 14° are shown. SEM images of polymer-functionalized platinum electrodes at a tilt angle of 14° are shown. SEM image of a non-functionalized platinum surface at a tilt angle of 1° is shown. SEM image of a non-functionalized platinum surface at a tilt angle of 45° is shown. SEM image of an unfunctionalized platinum surface at a tilt angle of 4° is shown. SEM image of a non-functionalized platinum surface at a tilt angle of 45° is shown.

DETAILED DESCRIPTION Provided herein are reagents, methods, and compositions for functionalizing or derivatizing metal surfaces. In some embodiments, the reagent comprises a phosphonic acid or phosphonic acid derivative (eg, a phosphonate ester). In some embodiments, metal surfaces include platinum and gold surfaces, or oxides thereof. In some embodiments, the metal surface is a platinum surface or its oxide. In some embodiments, the metal surface is a gold surface or its oxide. In some embodiments, the metal surface is a palladium, iridium, or rhodium surface, or oxides thereof. In some embodiments, the metal surface is a palladium surface or its oxide. In some embodiments, the metal surface is an iridium surface or its oxide. In some embodiments, the metal surface is a rhodium surface or its oxide. In some embodiments, the phosphonic acid or derivative thereof contains the desired surface-modifying groups. In some embodiments, the phosphonic acid or derivative thereof comprises a reactive handle for attachment to the desired surface modification group after modification with the phosphonic acid or derivative thereof. In some embodiments, the phosphonic acid or derivative thereof comprises a capture moiety for binding another group such as a biological binding moiety (eg, peptide or oligonucleotide). An example of a metal surface modified according to the invention is shown in FIG. FIG. 1A shows exemplary reactants and corresponding products bound to a metal surface (eg, a platinum surface). In some embodiments, the phosphonate reagent forms a metal phosphonate bond with the metal surface and has one of the binding modes shown in FIG. 1B. As detailed in FIG. 1B, the P=O bond of the phosphonate group is the interaction between the P=O oxygen and the metal atom on the metal surface, or the P=O oxygen and the adjacent -OH It can coordinate to the metal surface in various modes, such as interactions with groups (eg, when the metal surface is oxidized).

Also provided herein are uses of the modified metal surfaces provided herein. In some preferred embodiments, the modified metal surfaces are useful as antifouling surfaces for electrodes in electrophoresis-based assays. In some embodiments, these antifouling surfaces employ hydrogel layers. In some embodiments, the modified metal surfaces are useful for implantation into living organisms. In some embodiments, the modified metal surface has anti-corrosion properties compared to the unmodified metal surface. In some embodiments, modified metal surfaces are useful for creating active surfaces capable of binding analytes (eg, in biological capture assays).

Also provided herein are methods of making the modified metal surfaces described herein.

I. Modified Metal Surfaces In one aspect, provided herein are functionalized metal surfaces. In some embodiments, the metal surface is oxidized. In some embodiments, the metal surface comprises platinum or gold, or oxides thereof, or combinations thereof. In some embodiments, the metal surface comprises platinum or its oxide. In some embodiments, the metal surface comprises gold or its oxide. In some embodiments, the metal surface comprises palladium, iridium, or rhodium, or oxides thereof. In some embodiments, the metal surface comprises palladium or its oxide. In some embodiments, the metal surface comprises iridium or its oxide. In some embodiments, the metal surface comprises rhodium or its oxide. In some embodiments, the metal surface is a platinum or gold surface, or oxides thereof. In some embodiments, the metal surface is a platinum surface or its oxide. In some embodiments, the metal surface is a gold surface or its oxide. In some embodiments, the metal surface is palladium, iridium, or rhodium, or oxides thereof. In some embodiments, the metal surface is a palladium surface or its oxide. In some embodiments, the metal surface is an iridium surface or its oxide. In some embodiments, the metal surface is a rhodium surface or its oxide. In some embodiments, modified metal surfaces are attached to phosphonate moieties. In some embodiments, the phosphonate moiety is attached directly to the metal surface. In some embodiments, the phosphonate moiety forms a bond with the metal surface. In some embodiments, the phosphonate moiety forms a covalent bond with the metal surface.

In some embodiments, the metal surface is bound to phosphonate moieties. In some embodiments, the phosphonate moiety is a metal phosphonate. In some embodiments, surface-bound phosphonate moieties are prepared from phosphonate reagents or phosphonate derivatives. In some embodiments, the phosphonic acid derivative is a phosphonate ester. In some embodiments, the phosphonate ester is a C ₁ -C ₆ alkyl phosphonate ester. In some embodiments, the phosphonate ester is a methyl or ethyl phosphonate ester. In some embodiments, the phosphonic acid or phosphonate ester reagent is a bisphosphonic acid or bisphosphonate (eg, 1,1 bisphosphonic acid).

In some embodiments, the structure
Provided herein are functionalized metal surfaces comprising metal surfaces bound to molecules having In some embodiments, the functionalized metal surface comprises a metal surface bound to molecules as shown in FIG. 1B. In some embodiments, each
indicates the point of attachment to the metal surface. In some embodiments, one or more
is the point of attachment to the metal surface. In some embodiments, each
is the point of attachment to the metal surface. In some embodiments, described herein are R moieties attached to metal surfaces via phosphonate residues. In some embodiments, the R moiety is covalently attached to the metal surface via a phosphonate residue. In some embodiments, the metal surface comprises platinum or its oxide. In some embodiments, the metal surface comprises gold or its oxide. In some embodiments, the metal surface comprises palladium or its oxide. In some embodiments, the metal surface comprises iridium or its oxide. In some embodiments, the metal surface comprises rhodium or its oxide. In some embodiments, the metal surface is oxidized.

R in the formulas shown above can be any group. In some embodiments, R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted is optionally substituted alkynyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, R is optionally substituted alkyl or optionally substituted heteroalkyl.

In some embodiments, R directly contains the desired surface modification. In some embodiments, R is a hydrophobic residue, hydrophilic residue, charged residue, cationic residue, anionic residue, polysaccharide, hydrophobic polymer, hydrophilic polymer, antimicrobial agent, biomaterial , biocompatible materials, anti-fouling materials, conductive materials, semiconductor materials, heat-resistant materials, anti-corrosion materials, catalysts, or magnetic materials, or any combination thereof. In some embodiments, R comprises an oligonucleotide or polypeptide. In some embodiments, R comprises hydrogel.

Linking Moiety In some embodiments, the surface-modifying group is linked to the phosphonate group via a linking moiety. The linking moiety can be any suitable group capable of providing attachment of the surface modifying group to the phosphonate moiety.

In some embodiments, the linking moiety has the structure -L-, where L comprises an optionally substituted alkylene chain or an optionally substituted heteroalkylene chain. In some embodiments, the linking moiety comprises an optionally substituted alkylene chain. In some embodiments, the linking moiety comprises an unsubstituted alkylene chain. In some embodiments, the linking moiety comprises an optionally substituted heteroalkylene chain. In some embodiments, the linking moiety comprises an unsubstituted heteroalkylene chain. In some embodiments, a linking moiety comprises a chain of 1-100 atoms. In some embodiments, a linking moiety comprises a chain of 1-50 atoms. In some embodiments, a linking moiety comprises a chain of 1-40 atoms. In some embodiments, a linking moiety comprises a chain of 1-20 atoms. In some embodiments, the linking moiety comprises a chain of 5-100 atoms. In some embodiments, a linking moiety comprises a chain of 5-50 atoms. In some embodiments, the linking moiety comprises a chain of 5-40 atoms. In some embodiments, the linking moiety comprises a chain of 5-20 atoms. In some embodiments, the linking moiety comprises a chain of 10-100 atoms. In some embodiments, the linking moiety comprises a chain of 10-50 atoms. In some embodiments, the linking moiety comprises a chain of 10-40 atoms. In some embodiments, the linking moiety comprises a chain of 10-20 atoms.

In some embodiments, each atom in the chain is independently selected from C, N, O, S, Si, and P. In some embodiments, each atom in the chain is independently selected from C, N, O, and S. In some embodiments, each atom in the chain is independently selected from C, N, and O. In some embodiments, the first atom in the chain is C.

In some embodiments, L is of the formula -[Z-(CR ¹ R ² ] _n ] _m -
has the structure of
wherein each R ¹ and R ² is independently H or optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted or optionally substituted heteroaryl, or R ¹ and R ² on the same atom taken together form oxo, or R ¹ on adjacent atoms taken together forming a double bond or R ¹ and R ² on adjacent atoms together form a triple bond,
each Z is independently absent, -O-, -NR ³ -, -S-, -SS-, -S(O)-, -S(O) ₂ -, -C(O)-, - C(O)O—, —C(O)NR ³ , —OC(O)O—, —NR ³ C(O)NR ³ —, —OC(O)NR ³ —, —S(O) ₂ O -, -S(O) ₂ NR ³ -, -OS(O) ₂ O-, -NR ³ SO ₂ NR ³ -, or -OSO ₂ NR ³ -;
Each R ³ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl,
Each n is independently an integer from 1-30, and m is an integer from 1-30.

In some embodiments, each R ¹ and R ² is independently H, optionally substituted alkyl, optionally substituted heteroalkyl, or R ¹ and R ² together on the same atom together to form oxo, or R1 on adjacent atoms together form a double bond, or ^R1 and ^R2 on adjacent atoms together form a triple bond. In some embodiments, each R ¹ and R ² is independently H, optionally substituted alkyl, R ¹ and R ² on the same atom taken together form oxo, or R ¹ on adjacent atoms together form a double bond. In some embodiments, each R ¹ and R ² is independently H, optionally substituted alkyl, and R ¹ and R ² on the same atom together form oxo, or or R ¹ on adjacent atoms together form a double bond. In some embodiments, each R ¹ and R ² is independently H, optionally substituted alkyl, or R ¹ and R ² on the same atom together form oxo . In some embodiments, each R ¹ and R ² is independently H, optionally substituted C ₁ -C ₁₀ alkyl, or R ¹ and R ² taken together on the same atom to form oxo. In some embodiments, each R ¹ and R ² is independently H, optionally substituted alkyl, or R ¹ and R ² on the same atom together form oxo . In some embodiments, each R ¹ and R ² is independently C ₁ -C ₁₀ alkyl optionally substituted with H, hydroxy or alkoxy, or R ¹ and R ² on the same atom are Together they form oxo. In some embodiments, each R ¹ and R ² is independently H or R ¹ and R ² on the same atom together form oxo. In some embodiments, each R ¹ and R ² is H.

In some embodiments, each Z is independently absent, —O—, —NR ³ —, —S—, —C(O)—, —C(O)O—, —C( O)NR ³ , —OC(O)O—, —NR ³ C(O)NR ³ —, or —OC(O)NR ³ —. In some embodiments, each Z is independent. In some embodiments, each Z is independently absent, -O-, -NR ³ -, -C(O)O-, or -C(O)NR ³ . In some embodiments, each Z is independently absent, -O-, -C(O)O-, or -C(O) ^NR3 . In some embodiments, each Z is independently -O-, -C(O)O-, or -C(O) ^NR3 . In some embodiments, each Z is -O-. In some embodiments, each Z is -C(O)O- or -C(O) ^NR3 . In some embodiments, each Z is -C(O) ^NR3 .

In some embodiments, each R ³ is independently H, optionally substituted alkyl, or optionally substituted cycloalkyl. In some embodiments, each ^R3 is independently H or optionally substituted alkyl. In some embodiments, each R ³ is independently H or C ₁ -C ₆ alkyl optionally substituted with hydroxy or alkoxy. In some embodiments, each ^R3 is independently H or methyl.

In some embodiments, each n is independently an integer from 1-20. In some embodiments, each n is independently an integer from 1-15. In some embodiments, each n is independently an integer from 1-10. In some embodiments, each n is independently an integer from 2-20. In some embodiments, each n is independently an integer from 2-15. In some embodiments, each n is independently an integer from 2-10.

In some embodiments, m is an integer from 1-25. In some embodiments, m is an integer from 1-20. In some embodiments, m is an integer from 1-15. In some embodiments, m is an integer from 1-10. In some embodiments, m is an integer from 1-5. In some embodiments, m is an integer from 2-30. In some embodiments, m is an integer from 2-25. In some embodiments, m is an integer from 2-20. In some embodiments, m is an integer from 2-15. In some embodiments, m is an integer from 2-10. In some embodiments, m is an integer from 2-5.

L may contain one or more subunit building blocks. In some embodiments, L is
wherein each n is independently an integer from 1-30. In some embodiments, L is
comprising one or more subunits selected from In some embodiments, L is
comprising one or more subunits selected from In some embodiments, L is
comprising one or more subunits selected from

In some embodiments, L is the structure
wherein each n is independently an integer from 1-30 and each p is independently an integer from 1-30. In some embodiments, L is the structure
wherein each n is independently an integer from 1-30 and each p is independently an integer from 1-30. In some embodiments, L is the structure
have In some embodiments, L is the structure
have

In some embodiments, p is an integer from 1-20. In some embodiments, p is an integer from 1-15. In some embodiments, p is an integer from 1-10. In some embodiments, p is an integer from 1-5. In some embodiments, p is an integer from 2-30. In some embodiments, p is an integer from 2-20. In some embodiments, p is an integer from 2-15. In some embodiments, p is an integer from 2-10. In some embodiments, p is an integer from 2-5.

In some embodiments, L comprises a polymer. Any polymer can be used for the linking moiety L. Polymers may be branched or linear. In some embodiments, the polymer is polyethylene, polypropylene, polyvinyl halide, polystyrene, nylon, polyamide, polyester, polyaramid, polyacrylate, polymethacrylate, polytetrafluoroethylene, polysaccharide, poly(alkylene oxide), poly(vinylpyrrolidone ), poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine), or any combination or derivative thereof. In some embodiments, the polymer is polyethylene, nylon, polyamide, polyester, polyacrylate, polymethacrylate, poly(alkylene oxide), poly(vinylpyrrolidone), poly(vinyl alcohol), or any combination or derivative thereof. include. In some embodiments, the polymer comprises polyamides, polyesters, polyacrylates, polymethacrylates, poly(alkylene oxides), or any combination or derivative thereof. In some embodiments, the polymer comprises poly(alkylene oxide). In some embodiments the polymer comprises polyethylene glycol. In some embodiments the polymer comprises polymethacrylate. In some embodiments the polymethacrylate is a hydroxyalkyl methacrylate. In some embodiments, the polymethacrylate is a _C1 - _C20 hydroxyalkyl methacrylate. In some embodiments, the polymethacrylate is hydroxymethyl methacrylate, hydroxyethyl methacrylate, hydroxypropyl methacrylate, hydroxybutyl methacrylate, hydroxypentyl methacrylate, hydroxyhexyl methacrylate, hydroxyheptyl methacrylate, or hydroxyoctyl methacrylate.

In some embodiments, L comprises a cleavable linker. In some embodiments, the linking moiety comprises a cleavable linker. Any cleavable linker can be used in the present invention. In some embodiments, the cleavable linker is a photocleavable linker, a chemically cleavable linker, or an enzymatically cleavable linker.

Reactive Handles and Capture Moieties Linking moieties can be used to link phosphonate moieties to reactive handles. Reactive handles are configured to allow the addition of additional surface functionalizing reagents. A CLICK chemistry reagent is an example of a compatible reaction handle. Any suitable reactive group can be used. In some embodiments, the reactive handle is represented by X. In some embodiments, R has the structure XL-, where L is any of the linking moieties provided herein.

In some embodiments, X comprises a reactive functional group. Any suitable functional group can be used for this purpose. In some embodiments, X is an azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, acid, activated ester, alkene, alcohol, halide, acyl halide, sulfonic acid, sulfinic acid, including reactive functional groups selected from sulfonyl halides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, and hydrazides, or any combination thereof. In some embodiments, X is a reactive functional group selected from azides, alkynes, sulfhydryls, maleimides, activated esters, halides, acyl halides, epoxides, aldehydes, and ketones, or any combination thereof including. In some embodiments, X comprises reactive functional groups selected from azides, alkynes, sulfhydryls, maleimides, halides, and epoxides, or any combination thereof. In some embodiments, X includes azide. In some embodiments, X comprises alkyne. In some embodiments, X comprises cyclooctyne. In some embodiments, X comprises dibenzolcyclooctyne. In some embodiments, X comprises maleimide. In some embodiments, X comprises a halide. In some embodiments, X comprises an epoxide. In some embodiments, X comprises a sulfhydryl. In some embodiments, X comprises a carboxylic acid. In some embodiments, X comprises an amine. In some embodiments, X comprises an alkene. In some embodiments, X comprises a CLICK chemical reagent.

In some embodiments, X has the structure X ¹ -L ^X -Z ^X -, wherein:
Z ^X is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)-, -C(O)O -, -C(O)NR ⁴ , -OC(O)O-, -NR ⁴ C(O)NR ⁴ -, -OC(O)NR ⁴ -, -S(O) ₂ O-, -S( O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, or -OS(O) ₂ NR ⁴ -;
L ^X is absent, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl , optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted arylheteroalkyl, or optionally substituted heteroarylheteroalkyl;
X ¹ is an azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, carboxylic acid, active ester, alkene, alcohol, acyl halide, sulfonic acid, sulfinic acid, sulfonyl halide, epoxide, aldehyde, ketone , imines, oximes, isocyanates, isothiocyanates, hydrazines, or hydrazides, and each ^R4 is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl , optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

In some embodiments, ^ZX functions as a point of attachment linking the reactive handle to the linking moiety L. Any group that allows attachment of the reactive moiety to the linking moiety can be used. In some embodiments, Z ^X is a bond, —O—, —NR ⁴ —, —S—, —SS—, —C(O)—, —C(O)O—, —C(O) NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, or —OC(O)NR ⁴ —. In some embodiments, Z ^X is a bond, -NR ⁴ -, -O-, -C(O)-, -C(O)O-, or -C(O)NR ⁴ . In some embodiments, Z ^X is a bond. In some embodiments, Z ^X is -O-. In some embodiments, Z ^X is -NR ⁴ -. In some embodiments, Z ^X is -C(O)-. In some embodiments, Z ^X is -C(O)O-. In some embodiments, Z ^X is -C(O)NR ⁴ -.

^LX functions as a secondary linking moiety between the point of attachment to linking moiety L and the reactive functional group of X. Any suitable linker can be used. In some embodiments, L ^X is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted hetero alkylene, optionally substituted arylene, optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene. In some embodiments, L ^X is optionally substituted alkylene or optionally substituted heteroalkylene. In some embodiments, L ^X is optionally substituted alkylene. In some embodiments, L ^X is alkylene. In some embodiments, L ^X is an optionally substituted heteroalkylene. In some embodiments, L ^X is heteroalkylene. In some embodiments, ^LX comprises a cleavable linker group.

In some embodiments, L ^X has the structure -(CR ⁵ R ⁶ ) _r -
has
wherein each ^R5 and ^R6 is independently H, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl, or ^R5 and ^R6 taken together on the same atom, oxo, or ^R5 on adjacent atoms taken together, a double bond or adjacent atoms R ⁵ and R ⁶ together form a triple bond, and r is an integer from 0-30.

In some embodiments, R ⁵ and R ⁶ are each independently H, optionally substituted alkyl, or optionally substituted heteroalkyl, or R ⁵ and R ⁶ on the same atom are Together form oxo, or ^R5s of adjacent atoms together form a double bond. In some embodiments, R ⁵ and R ⁶ are each independently H, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each ^R5 and ^R6 is independently H, optionally substituted alkyl. In some embodiments, each R ⁵ and R ⁶ is independently H or optionally substituted C ₁ -C ₆ alkyl. In some embodiments, each R ⁵ and R ⁶ is independently H, or C ₁ -C ₆ alkyl. In some embodiments, each R ⁵ and R ⁶ is independently H, or C ₁ -C ₃ alkyl.

In some embodiments, r is an integer from 0-30. In some embodiments, r is an integer from 0-20. In some embodiments, r is an integer from 0-10. In some embodiments, r is an integer from 0-5. In some embodiments, r is 0 or 1.

In some embodiments, X ¹ is an azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, carboxylic acid, activated ester, alkene, alcohol, acyl halide, sulfonic acid, sulfinic acid, halogen sulfonyl chlorides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, or hydrazides. In some embodiments, X ¹ is an azide, alkyne, halide, sulfhydryl, maleimide, amine, carboxylic acid, alkene, alcohol, epoxide, aldehyde, ketone, imine, or hydrazine. In some embodiments, X ¹ is an amine, carboxylic acid, azide, alkyne, halide, alkene, or sulfhydryl.

In some embodiments, X is the structure
have In some embodiments, X is the structure
have In some embodiments, X is the structure
have In some embodiments, X is the structure
have

In some embodiments, X is
is. In some embodiments, X is
is. In some embodiments, X is
is. In some embodiments, X is
is. In some embodiments, X is
is. In some embodiments, X is
is.

X can also include a trapping moiety. A capture moiety can be any type of molecule capable of selectively capturing a desired substance. In some embodiments, capture moieties include biotin, avidin, streptavidin, nucleic acids, peptides, proteins, or any combination thereof. In some embodiments the capture moiety comprises biotin. In some embodiments, the capture moiety comprises avidin. In some embodiments, the capture moiety comprises streptavidin. In some embodiments, capture moieties comprise nucleic acids. In some embodiments, the nucleic acid is an aptamer. In some embodiments the nucleic acid is a capture probe. In some embodiments the capture moiety is a peptide. In some embodiments, the capture moieties are proteins.

Any of the modified metal surfaces provided herein can be manufactured or distributed as kits. In some embodiments, the kit includes a modified metal surface and instructions for use. In some embodiments, the modified metal surface comprises reactive handles or capture moieties attached to the metal surface. In some embodiments, the instructions provide protocols for linking additional moieties to the metal surface via reactive handles or capture moieties.

Surface Modifications In some embodiments, surface modifications are attached to the phosphonate moiety by a linking moiety. In some embodiments, the surface modification is attached directly to the phosphonate moiety. In some embodiments, the surface modifying group is attached to the phosphonate moiety via the reaction product of either the reactive handle or the capture moiety X described above.

Any desired surface modifying group can be applied to the metal surface by the metal phosphonate linkages described herein. In some embodiments, the surface modifying groups are hydrophobic residues, hydrophilic residues, charged residues, cationic residues, anionic residues, polysaccharides, hydrophobic polymers, hydrophilic polymers, antimicrobial agents, selected from biomaterials, biocompatible materials, antifouling materials, conductive materials, semiconductor materials, heat resistant materials, anticorrosion materials, catalysts, and magnetic materials, or any combination thereof. In some embodiments the surface-modifying group is a hydrogel.

In some embodiments, the surface modification is represented by Y. In some embodiments, R has the structure YL-, where L is any of the linking moieties provided herein.

In some embodiments, Y has the structure Y ¹ -L ^Y -Z ^Y -. In some embodiments Y ¹ represents a surface modifying group. In some embodiments, ^LY serves as a secondary linking moiety between the point of attachment to linking moiety L and the surface modifying group of Y. In some embodiments, ^ZY serves as the point of attachment linking the surface-modifying group to the linking moiety L.

In some embodiments, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)- , —C(O)O—, —C(O)NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O-, -S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -, or azide, alkyne , tetrazines, halides, sulfhydryls, disulfides, maleimides, amines, carboxylic acids, active esters, alkenes, alcohols, acyl halides, sulfonic acids, sulfinic acids, sulfonyl halides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, iso A reaction product formed between a thiocyanate, hydrazine, or hydrazide and a suitable complementary reactive group. In some embodiments, Z ^Y is azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, carboxylic acid, activated ester, alkene, alcohol, acyl halide, sulfonic acid, sulfinic acid, halogen It is a reaction product formed between a sulfonyl group, epoxide, aldehyde, ketone, imine, hydrazine, or hydrazide and a suitable complementary reactive group. In some embodiments, Z ^Y is a reaction product formed by a covalent bond-forming reaction between an azide, alkyne, halide, sulfhydryl, maleimide, alkene, or epoxide and a suitable complementary reactive group. . In some embodiments, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)- , —C(O)O—, —C(O)NR ⁴ , —OC(O)O—, —NR ₄ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O-, -S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -. In some embodiments, Z ^Y is a bond, —O—, —NR ⁴ —, —S—, —SS—, —C(O)—, —C(O)O—, or —C(O ) NR ⁴ . In some embodiments, ZY is -SS-, -C(O)-, -C(O)O-, or -C(O) ^NR4 . In some embodiments, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)- , —C(O)O—, —C(O)NR ⁴ —, —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O ) ₂ O-, -S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -,
is. In some embodiments, Z ^Y is
is. In some embodiments, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)- , —C(O)O—, —C(O)NR ⁴ —, —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O ) ₂ O-, -S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -.

In some embodiments, L ^Y is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted hetero alkylene, optionally substituted arylene, optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene. In some embodiments, L ^Y is absent, optionally substituted alkylene, or optionally substituted heteroalkylene. In some embodiments, L ^Y is alkylene. In some embodiments, L ^Y is heteroalkylene. In some embodiments, ^LY is absent. In some embodiments, ^LY includes a cleavable linker group.

In some embodiments, each R ⁴ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, each R ⁴ is independently H, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each R ⁴ is independently H or C ₁ -C ₆ alkyl. In some embodiments, each ^R4 is independently H or methyl.

In some embodiments, Y ¹ comprises at least one surface modification residue. Any suitable surface modification residue can be used.

In some embodiments, Y ¹ comprises a hydrophobic residue. In some embodiments, the hydrophobic residue is a fatty acid or fatty acid derivative. In some embodiments the hydrophobic residue is a fluorinated fatty acid.

In some embodiments, the hydrophobic residue comprises a hydrophobic polymer. In some embodiments, the hydrophobic polymer comprises polyethylene, polypropylene, polystyrene, polyvinyl halide, polytetrafluoroethylene, polymethylmethacrylate, polycarbonate, polyetherurethane, polydimethylsiloxane, or any combination or derivative thereof. . In some embodiments the hydrophobic polymer comprises polyethylene. In some embodiments the hydrophobic polymer comprises polypropylene. In some embodiments the hydrophobic polymer comprises polystyrene. In some embodiments, the hydrophobic polymer comprises polyvinyl halide. In some embodiments the hydrophobic polymer comprises polytetrafluoroethylene. In some embodiments, the hydrophobic polymer comprises polymethylmethacrylate. In some embodiments, the hydrophobic polymer comprises polycarbonate. In some embodiments, the hydrophobic polymer comprises polyetherurethane. In some embodiments, the hydrophobic polymer comprises polydimethylsiloxane. In some embodiments, the hydrophobic polymer comprises polysilane. In some embodiments the hydrophobic polymer comprises a fluoropolymer.

In some embodiments, Y ¹ comprises a hydrophilic residue. Any suitable hydrophilic residue can be used. In some embodiments, hydrophilic residues comprise monosaccharides or polysaccharides. In some embodiments, hydrophilic residues comprise hydrophilic polymers. In some embodiments, the hydrophilic polymer is polyacrylamide, polyacrylate, polymethacrylate, poly(alkylene oxide), poly(vinylpyrrolidone), poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine). or any combination of derivatives thereof. In some embodiments, the hydrophilic polymer comprises polyacrylamide. In some embodiments, the hydrophilic polymer comprises polyacrylate. In some embodiments, the hydrophilic polymer comprises polymethacrylate. In some embodiments, the hydrophilic polymer comprises poly(alkylene oxide). In some embodiments, the hydrophilic polymer comprises poly(vinylpyrrolidone). In some embodiments, the hydrophilic polymer comprises poly(vinyl alcohol). In some embodiments, hydrophilic polymers include polyoxazolines. In some embodiments, the hydrophilic polymer comprises poly(N-acryloylmorpholine).

In some embodiments, the hydrophilic polymer is polyacrylamide, poly(diethylacrylamide), poly(dimethylacrylamide), poly(N-isopropylacrylamide), poly(acrylic acid), poly(methacrylic acid), poly(acrylamide acid methyl), poly(ethyl acrylate), poly(2-hydroxyethyl acrylate), poly(propyl acrylate), poly(butyl acrylate), poly(methyl methacrylate), poly(2-hydroxyethyl methacrylate), poly(tetrahydrofuran furyl methacrylate), poly(ethylene oxide), poly(propylene oxide), poly(vinylpyrrolidone), polyoxazoline, poly(2-ethyloxazoline), or poly(N-acryloylmorpholine). In some embodiments, the hydrophilic polymer comprises polyacrylamide. In some embodiments, the hydrophilic polymer comprises poly(N,N-diethylacrylamide). In some embodiments, the hydrophilic polymer comprises poly(dimethylacrylamide). In some embodiments, the hydrophilic polymer comprises poly(N-isopropylacrylamide). In some embodiments, the hydrophilic polymer comprises poly(acrylic acid). In some embodiments, the hydrophilic polymer comprises poly(methacrylic acid). In some embodiments, the hydrophilic polymer comprises poly(methyl acrylate). In some embodiments, the hydrophilic polymer comprises poly(ethyl acrylate). In some embodiments, the hydrophilic polymer comprises poly(2-hydroxyethyl acrylate). In some embodiments, the hydrophilic polymer comprises poly(propyl acrylate). In some embodiments, the hydrophilic polymer comprises poly(butyl acrylate). In some embodiments, the hydrophilic polymer comprises poly(methyl methacrylate). In some embodiments, the hydrophilic polymer comprises polymethacrylate. In some embodiments the polymethacrylate is a hydroxyalkyl methacrylate. In some embodiments, the polymethacrylate is a _C1 - _C20 hydroxyalkyl methacrylate. In some embodiments, the polymethacrylate is hydroxymethyl methacrylate, hydroxyethyl methacrylate, hydroxypropyl methacrylate, hydroxybutyl methacrylate, hydroxypentyl methacrylate, hydroxyhexyl methacrylate, hydroxyheptyl methacrylate, or hydroxyoctyl methacrylate. In some embodiments, the hydrophilic polymer comprises poly(2-hydroxyethyl methacrylate). In some embodiments, the hydrophilic polymer comprises poly(tetrahydrofurfuryl methacrylate). In some embodiments, the hydrophilic polymer comprises poly(ethylene oxide). In some embodiments, the hydrophilic polymer comprises poly(propylene oxide). In some embodiments, the hydrophilic polymer comprises poly(vinylpyrrolidone). In some embodiments, hydrophilic polymers include polyoxazolines. In some embodiments, the hydrophilic polymer comprises poly(2-ethyloxazoline). In some embodiments, the hydrophilic polymer comprises poly(N-acryloylmorpholine).

In some embodiments, the surface-modifying group of Y has the structure
including
During the ceremony,
each R ⁷ is independently optionally substituted C ₁ -C ₃ alkyl or optionally substituted C ₁ -C ₃ hydroxyalkyl;
each R ⁸ is independently H or C ₁ -C ₃ alkyl;
each X ⁷ is independently absent, —O—, —S—, or NR9;
Each R ⁹ is independently H or optionally substituted C ₁ -C ₆ alkyl, and s is an integer from 1-1,000,000.

In some embodiments, the structure
is connected to the connecting part.

In some embodiments, each R ⁷ is independently H, C ₁ -C ₃ alkyl or C ₁ -C ₃ hydroxyalkyl. In some embodiments, each R ⁷ is independently H, methyl, ethyl, propyl, isopropyl, 2-hydroxyethyl, 3-hydroxypropyl, or 2-hydroxypropyl.

In some embodiments, each ^R8 is independently H, ethyl, methyl, or propyl. In some embodiments, each ^R8 is independently H or methyl. In some embodiments, each ^R8 is H. In some embodiments, each ^R8 is methyl.

In some embodiments, each X ⁷ is independently absent, -O-, -S-, or NR ⁹ . In some embodiments, each X ⁷ is independently -O- or -NR ⁹ . In some embodiments, each X ⁷ is -O-. In some embodiments, each X ⁷ is independently -NR ⁹ . In some embodiments, each ^X7 is absent.

In some embodiments, each R ⁹ is independently H or C ₁ -C ₃ alkyl optionally substituted with hydroxy or alkoxy. In some embodiments, each R ⁹ is independently H or C ₁ -C ₃ alkyl. In some embodiments, each ^R9 is independently H or methyl. In some embodiments, each ^R9 is H.

In some embodiments, s is an integer from 1-1,000,000. In some embodiments, s is an integer from 1-100,000. In some embodiments, s is an integer from 1-10,000. In some embodiments, s is an integer from 1-1000. In some embodiments, s is an integer from 1-100. In some embodiments, s is an integer from 10-1,000,000. In some embodiments, s is an integer from 10-100,000. In some embodiments, s is an integer from 10-10,000. In some embodiments, s is an integer from 10 to 1000. In some embodiments, s is an integer from 10-100. In some embodiments, s is an integer from 1-100. In some embodiments, s is an integer from 100-1,000,000. In some embodiments, s is an integer from 100 to 100,000. In some embodiments, s is an integer from 10-10,000. In some embodiments, s is an integer from 100-1000.

In some embodiments, surface-modifying groups comprise cationic residues. In some embodiments, Y ¹ comprises a cationic residue. In some embodiments, cationic residues comprise amine groups, substituted amine groups, quaternary amine groups, or any combination thereof. In some embodiments, the cationic residue is an amine group. In some embodiments, the cationic residue is a substituted amine group. In some embodiments, the cationic residue comprises protonated amine groups, protonated substituted amine groups, or quaternary amine groups, or any combination thereof.

In some embodiments, Y ¹ comprises a cationic polymer. Any polymerizable monomer containing cationic groups can be used. In some embodiments, the cationic polymer is poly(ethyleneimine) (PEI), polyamino acids (e.g., polylysine, polyarginine), polypyridinium, polyammonium, polyacrylates containing amino groups, poly Contains acrylamide or chitosan.

In some embodiments, surface-modifying groups comprise anionic residues. In some embodiments, Y ¹ comprises an anionic residue. Any anionic residue can be used. In some embodiments, anionic residues include carboxylates, sulfonates, sulfinates, phosphates, phosphonates, or any combination thereof. In some embodiments the anionic residue is a carboxylate. In some embodiments the anionic residue is a sulfonate. In some embodiments the anionic residue is a sulfinate. In some embodiments the anionic residue is a phosphate. In some embodiments the anionic residue is a phosphonate.

In some embodiments, surface-modifying groups comprise anionic polymers. In some embodiments, Y ¹ comprises an anionic polymer. In some embodiments, the anionic polymer comprises anionic groups selected from carboxylates, sulfonates, sulfinates, phosphates, and phosphonates, or any combination thereof. In some embodiments, anionic polymers comprise polyamino acids (eg, polyglutamic acid, polyaspartic acid), polyacrylates, polymethacrylates, polysulfonates, or polyacids.

In some embodiments, the surface-modifying groups are zwitterionic groups. In some embodiments, Y ¹ comprises a zwitterionic group. In some embodiments, Y comprises a zwitterionic group. Any combination of cationic or anionic groups listed herein can be used.

In some embodiments, surface-modifying groups comprise hydrogels. In some embodiments Y comprises a hydrogel. In some embodiments, Y ¹ comprises a hydrogel. In some embodiments, R comprises hydrogel. Any hydrogel can be linked with the phosphonate reagents herein. Hydrogel properties can be modified for any desired purpose of the functionalized metal surfaces provided herein. In some embodiments, hydrogel properties are optimized for use with metal electrodes. A detailed description of hydrogels that may adhere to metal surfaces can be found in the "Hydrogel Surface Modification" section below.

In some embodiments, surface-modifying groups comprise probe moieties. In some embodiments, Y comprises a probe moiety. In some embodiments, Y ¹ comprises a probe moiety. In some embodiments, R comprises a probe moiety. Any type of probe portion can be used. In some embodiments, the probe portion comprises an analyte or target specific binding agent. In some embodiments, probe moieties comprise nucleic acids, peptides, proteins, antibodies, small molecules, glycans, or any combination thereof.

In some embodiments, the probe portion comprises nucleic acids. In some embodiments, nucleic acids comprise DNA or RNA. In some embodiments, the nucleic acid is configured to hybridize with a target moiety or analyte. In some embodiments, nucleic acids include aptamers. In some embodiments, nucleic acids comprise non-naturally occurring oligonucleotides.

In some embodiments, probe moieties comprise peptides. In some embodiments, probe moieties comprise proteins. In some, probe moieties include protein fragments. In some embodiments, the probe portion comprises an antibody. In some embodiments, the probe portion comprises an antibody fragment. In some embodiments, probe moieties comprise small molecules. In some embodiments, probe moieties comprise glycans.

In some embodiments, surface-modifying groups comprise drug moieties. The drug moiety can be linked to the phosphonate moiety by any linking moiety. In some embodiments, the drug moiety is linked to the surface via a cleavable linker. In some embodiments, the linker is configured to release the drug moiety.

In some embodiments, the surface-modifying group comprises a targeting moiety. In some embodiments, targeting moieties are specific to a particular biomarker. A targeting moiety can be any suitable group. In some embodiments, targeting moieties comprise antibodies or antibody fragments, peptides, or nucleic acids. In some embodiments, the targeting moiety and drug moiety are attached to the same surface.

Hydrogel Surface Modifications Also provided herein are hydrogels that can be linked to metal surfaces via bonds between the phosphonate groups and the metal surface. In addition to being applied to platinum, gold, palladium, iridium, or rhodium surfaces as described herein, these hydrogels can also be used on silicon, aluminum, titanium, iron, zinc, zirconium, nickel, silver, copper , cobalt, or chromium, or their oxides, or any combination thereof, to other metal surfaces capable of forming bonds with phosphonates. In some embodiments, the hydrogel is attached to a metal surface comprising platinum. In some embodiments, the hydrogel is attached to a metal surface comprising gold. In some embodiments, the hydrogel is attached to a metal surface comprising palladium. In some embodiments, the hydrogel is attached to a metal surface comprising iridium. In some embodiments, the hydrogel is attached to a metal surface comprising rhodium. In some embodiments, the hydrogel is attached to a metal surface comprising silicon. In some embodiments, the hydrogel is attached to a metal surface comprising aluminum. In some embodiments, the hydrogel is attached to a metal surface comprising titanium. In some embodiments, the hydrogel is attached to a metal surface comprising iron. In some embodiments, the hydrogel is attached to a metal surface comprising zinc.

The hydrogels provided herein may be linked to a metal surface via any of the linking moieties provided herein, including the linking moieties provided in the "Connecting Moieties" section, or there may be linked via a bond formed by reaction of any of the reactive handles provided in the .

In some embodiments, surface-modifying groups comprise hydrogels. In some embodiments Y comprises a hydrogel. In some embodiments, Y ¹ comprises a hydrogel. In some embodiments, R comprises hydrogel. Any hydrogel can be linked with the phosphonate reagents herein. The properties of the hydrogel can be modified for any desired purpose of the functionalized metal surfaces provided herein. In some embodiments, hydrogel properties are optimized for use with metal electrodes.

In some embodiments, hydrogels are used on electrodes configured for electrophoretic analysis of biological samples. If the metal surface constitutes an electrode, covering the electrode structure with one or more layers of material may prevent electrolysis reactions, heating, and chaotic fluid movement that may occur on or near the electrode. Adverse electrochemical effects, including but not limited to, can be reduced while still allowing effective separation of cells, bacteria, viruses, nanoparticles, DNA, and other biomolecules. In some embodiments, the hydrogel laminated onto the electrode structure can include one or more porous layers.

In some embodiments, the hydrogel has sufficient mechanical strength and is relatively chemically inert to withstand electrochemical effects at the electrode surface without deformation or degradation. In some embodiments, the hydrogel is sufficiently permeable to small aqueous ions, but keeps biomolecules away from the electrode surface. In some embodiments, the hydrogel comprises a porosity gradient, with the bottom of the hydrogel layer having a greater porosity than the top of the hydrogel layer.

In some embodiments Y comprises a hydrogel. In some embodiments, Y ¹ comprises a hydrogel.

In some embodiments, the hydrogel has a conductivity of about 0.001 S/m to about 10 S/m. In some embodiments, hydrogels have a conductivity from about 0.01 S/m to about 10 S/m. In some embodiments, hydrogels have a conductivity from about 0.1 S/m to about 10 S/m. In some embodiments, the hydrogel has a conductivity of about 1.0 S/m to about 10 S/m. In some embodiments, hydrogels have a conductivity from about 0.01 S/m to about 5 S/m. In some embodiments, hydrogels have a conductivity from about 0.01 S/m to about 4 S/m. In some embodiments, the hydrogel has a conductivity of about 0.01 S/m to about 3 S/m. In some embodiments, hydrogels have a conductivity of about 0.01 S/m to about 2 S/m. In some embodiments, hydrogels have a conductivity from about 0.1 S/m to about 5 S/m. In some embodiments, the hydrogel has a conductivity of about 0.1 S/m to about 4 S/m. In some embodiments, hydrogels have a conductivity of about 0.1 S/m to about 3 S/m. In some embodiments, hydrogels have a conductivity of about 0.1 S/m to about 2 S/m. In some embodiments, hydrogels have a conductivity of about 0.1 S/m to about 1.5 S/m. In some embodiments, the hydrogel has a conductivity of about 0.1 S/m to about 1.0 S/m.

In some embodiments, the hydrogel has a conductivity of about 0.1 S/m. In some embodiments, hydrogels have a conductivity of about 0.2 S/m. In some embodiments, the hydrogel has a conductivity of about 0.3 S/m. In some embodiments, the hydrogel has a conductivity of about 0.4 S/m. In some embodiments, the hydrogel has a conductivity of about 0.5 S/m. In some embodiments, the hydrogel has a conductivity of about 0.6 S/m. In some embodiments, the hydrogel has a conductivity of about 0.7 S/m. In some embodiments, the hydrogel has a conductivity of about 0.8 S/m. In some embodiments, the hydrogel has a conductivity of about 0.9 S/m. In some embodiments, the hydrogel has a conductivity of about 1.0 S/m.

In some embodiments, hydrogels have a thickness of about 0.001 microns to about 10 microns. In some embodiments, the hydrogel has a thickness of about 0.001 microns to about 5 microns. In some embodiments, the hydrogel has a thickness of about 0.001 micron to about 1 micron. In some embodiments, the hydrogel has a thickness of about 0.001 microns to about 0.5 microns. In some embodiments, the hydrogel has a thickness of about 0.01 microns to about 10 microns. In some embodiments, the hydrogel has a thickness of about 0.01 microns to about 5 microns. In some embodiments, hydrogels have a thickness of about 0.01 to about 1 micron. In some embodiments, the hydrogel has a thickness of about 0.1 microns to about 10 microns. In some embodiments, the hydrogel has a thickness of about 0.1 microns to about 5 microns. In some embodiments, the hydrogel has a thickness of about 0.1 microns to about 4 microns. In some embodiments, the hydrogel has a thickness of about 0.1 microns to about 3 microns. In some embodiments, the hydrogel has a thickness of about 0.1 microns to about 2 microns. In some embodiments, the hydrogel has a thickness of about 1 micron to about 5 microns. In some embodiments, the hydrogel has a thickness of about 1 micron to about 4 microns. In some embodiments, the hydrogel has a thickness of about 1 micron to about 3 microns. In some embodiments, the hydrogel has a thickness of about 1 micron to about 2 microns. In some embodiments, hydrogels have a thickness of about 0.5 microns to about 1 micron.

In some embodiments, hydrogels comprise any suitable synthetic polymer that forms hydrogels. In general, any molecule that is sufficiently hydrophilic and polymerizable can be utilized in the preparation of synthetic polymer hydrogels for the uses disclosed herein. Polymerizable moieties in the monomer may include alkenyl moieties, in which a double bond is double bonded to an oxygen and a single bond is bonded to another oxygen, nitrogen, sulfur, halogen, or carbon. substituted or unsubstituted α, β, unsaturated carbonyls directly attached to the carbon that is attached to the carbon; vinyls in which the double bond is single-bonded to oxygen, nitrogen, halogen, phosphorus or sulfur; Allyl Single Bonded to Carbon Bonded to Oxygen, Nitrogen, Halogen, Phosphorus or Sulfur; Double Bond Single Bond to Carbon which is Single Bond to Oxygen, Nitrogen, Halogen, Phosphorus or Sulfur homoaryl, which is single bonded to another carbon atom; alkynyl moieties, where the triple bond resides between two carbon atoms. In some embodiments, acryloyl or acrylamide monomers such as acrylates, methacrylates, acrylamides, methacrylamides are useful for forming hydrogels disclosed herein. More preferred acrylamide monomers include acrylamide, N-substituted acrylamide, N-substituted methacrylamide, and methacrylamide. In some embodiments, the hydrogel comprises epoxide-based polymers, vinyl-based polymers, allyl-based polymers, homoallyl-based polymers, cyclic anhydride-based polymers, ester-based polymers, ether-based polymers, alkylene glycol Including polymers such as base (eg, polypropylene glycol) polymers.

In some embodiments, the hydrogel comprises poly(2-hydroxyethyl methacrylate) (pHEMA), cellulose acetate, cellulose acetate phthalate, cellulose acetate butyrate, or any suitable acrylamide or vinyl-based polymer, or derivative thereof. include.

In some embodiments, the hydrogel is polymerized by atom transfer radical polymerization (ATRP). In some embodiments, the hydrogel is polymerized by reversible addition-fragmentation chain transfer (RAFT) polymerization. In some embodiments, the hydrogel is polymerized onto a metal surface via a reactive group attached to a phosphonate attached to the metal surface, the reactive group comprising a halide alpha to the carbonyl. In some embodiments, the alpha position to the carbonyl is a tertiary carbon. In some embodiments, the reactive group has the structure (CH ₃ ) ₂ BrC--C(O)--.

In some embodiments, additives are added to the hydrogel to increase the electrical conductivity of the gel. In some embodiments, hydrogel additives are conductive polymers (e.g., PEDOT:PSS), salts (e.g., copper chloride), metals (e.g., gold), plasticizers (e.g., PEG200, PEG400, or PEG600). , or a co-solvent. . In some embodiments, hydrogels help maintain the stability of DNA hybrids including, but not limited to, histidine, histidine peptides, polyhistidine, lysine, lysine peptides, and other cationic compounds or substances. Also includes compounds or materials.

Metal Surfaces Any suitable metal surface can be modified with the compositions and systems according to the present disclosure. In some embodiments, the metal surface is oxidized. A metal surface modified according to the present disclosure can be of any shape so long as there is a metal surface available to react with the phosphonic acid or phosphonate ester reagents described herein. In some embodiments, the metal surface comprises platinum or its oxide. In some embodiments, the metal surface comprises gold or its oxide. In some embodiments, the metal surface comprises palladium or its oxide. In some embodiments, the metal comprises iridium or its oxide. In some embodiments, the metal surface comprises a rhodium surface or its oxide. In some embodiments, the metal surface is a platinum surface or its oxide. In some embodiments, the metal surface is a gold surface or its oxide. In some embodiments, the metal surface is a palladium surface or its oxide. In some embodiments, the metal surface is an iridium surface or its oxide. In some embodiments, the metal surface is a rhodium surface or its oxide. In some embodiments, the metal surface comprises silicon, aluminum, titanium, iron, zinc, zirconium, nickel, silver, copper, cobalt, or chromium. In some embodiments, the metal surface is oxidized.

In some embodiments, the metal surface is a flat metal surface. In some embodiments, the metal surface is a curved metal surface. In some embodiments, the metal surface has a 3D shape. In some embodiments, the metal surface is an electrode, microchip, bead, microparticle, or nanoparticle. In some embodiments, the metal surface is an electrode. In some embodiments, the metal surface is a microtip. In some embodiments, the metal surface is a bead. In some embodiments, the metal surface is particulate. In some embodiments, the metal surfaces are nanoparticles.

Any portion of a metal surface can be modified with the surface modifications provided herein. In some embodiments, at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the metal surface is functionalized. In some embodiments, up to about 1%, up to about 2%, up to about 5%, up to about 10%, up to about 20%, up to about 30%, up to about 40%, up to about 50%, up to about 60%, up to about 70%, up to about 80%, or up to about 90% of the metal surface is functionalized. In some embodiments, about 1%, about 2%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% , about 90%, or substantially all metal surfaces are functionalized. In some embodiments, the degree of surface functionalization is measured by the degree of coverage of the surface modifying material on the surface. In some embodiments, the degree of surface functionalization is measured by the amount of phosphonate residues attached to the surface.
In some embodiments, the metal surface is modified with phosphonate at a concentration of about 0.0001 nmol/cm ² to about 5 nmol/cm ² . In some embodiments, the metal surface is about 0.0001 nmol ^/ cm ² to about 0.001 nmol/cm 2 , about 0.0001 nmol/cm ² to about 0.01 nmol/cm ² , about 0.0001 nmol/cm ^{2 .} to about 0.05 nmol/cm ² , about 0.0001 nmol/cm ² to about 0.1 nmol/cm ² , about 0.0001 nmol/cm ² to about 0.5 nmol/cm ² , about 0.0001 nmol/cm ² to about 1 nmol/ ^cm2 , about 0.0001 nmol/ ^cm2 to about 1.5 nmol/ ^cm2 , about 0.0001 nmol/ ^cm2 to about 2 nmol/ ^cm2 , about 0.0001 nmol/ ^cm2 to about 5 nmol/ ^cm2 , about 0.001 nmol/cm ² to about 0.01 nmol/cm ² , about 0.001 nmol/cm ² to about 0.05 nmol/cm 2 , about ^0.001 nmol/cm ² to about 0.1 nmol/cm ² , about 0. 001 nmol/cm ² to about 0.5 nmol/cm ² , about 0.001 nmol/cm ² to about 1 nmol/cm 2 , ^about 0.001 nmol/cm ² to about 1.5 nmol/cm ² , about 0.001 nmol/cm ² to about 2 nmol/cm ² , from about 0.001 nmol/cm ² to about 5 nmol/cm ² , from about 0.01 nmol/cm ² to about 0.05 nmol/cm ² , from about 0.01 nmol/cm ² to about 0.1 nmol/cm 2 cm ² , about 0.01 nmol/cm ² to about 0.5 nmol/cm ² , about 0.01 nmol/cm ² to about 1 nmol/cm ² , about 0.01 nmol/cm ² to about 1.5 nmol/cm ² , about 0.01 nmol/cm ² to about 2 nmol/cm ² , about 0.01 nmol/cm ² to about 5 nmol/cm ² , about 0.05 nmol/cm ² to about 0.1 nmol/cm ² , about 0.05 nmol/cm ² to about 0.5 nmol/cm ² , from about 0.05 nmol/cm ² to about 1 nmol/cm ² , from about 0.05 nmol/cm ² to about 1.5 nmol/cm ² , from about 0.05 nmol/cm ² to about 2 nmol/cm 2 cm ² cm ² , about 0.05 nmol/cm ² to about 5 nmol/cm ² , about 0.1 nmol/cm ² to about 0.5 nmol/cm 2 , about 0.1 nmol/ ^{cm 2 to} about 1 nmol/cm ² , about 0.1 nmol/cm ² to about 1.5 nmol/cm ² , about 0.1 nmol/cm ² to about 2 nmol/cm ² , about 0.1 nmol/cm ² to about 5 nmol/cm ² , about 0.5 nmol/cm ² to about 1 nmol/cm ² , from about 0.5 nmol/cm ² to about 1.5 nmol/cm ^{2 , from about 0.5 nmol/cm 2 to} about 2 nmol/cm ² , from about 0.5 nmol/cm ² to about 5 nmol/cm ² , from about 1 nmol/cm ² to about 1.5 nmol/cm ² , from about 1 nmol/cm ² to about 2 nmol/cm ² , from about 1 nmol/cm ² to about 5 nmol/cm ² , from about 1.5 nmol/cm ² to about 2 nmol/cm 2 . cm ² , from about 1.5 nmol/cm ² to about 5 nmol/cm ² , or from about 2 nmol/cm ² to about 5 nmol/cm ² . In some embodiments, the metal surface is about 0.0001 nmol/cm ² , about 0.001 nmol/cm 2 , about 0.01 nmol/cm ² , about 0.05 nmol/cm ² , about 0.1 nmol ^/ cm ² . ^{. _ _ _ _} In some embodiments, ^the metal surface is at least about 0.0001 nmol/cm ² , about 0.001 nmol/cm 2 , about 0.01 nmol/cm ² , about 0.05 nmol/cm ² , about 0.1 nmol/cm 2 . ² , modified with phosphonate at a concentration of about 0.5 nmol/cm ² , about 1 nmol/cm ² , about 1.5 nmol/cm ² , or about 2 nmol/cm ² . In some embodiments, the metal surface is at most about 0.001 nmol/cm ² , about 0.01 nmol/cm ² , about 0.05 nmol/cm 2 , about 0.1 nmol/ ^{cm 2 ,} about 0.5 nmol/cm 2 . cm ² , about 1 nmol/cm ² , about 1.5 nmol/cm ² , about 2 nmol/cm ² , or about 5 nmol/cm ² . In some embodiments, the metal surface is modified with phosphonate at a concentration of about 0.01 nmol/cm ² to about 2 nmol/cm ² .

In some embodiments, the metal surface is oxidized. In some embodiments, the metal surface is oxidized to allow attachment of the phosphonate moieties of the surface modifications provided herein. In some embodiments, the metal surface is partially oxidized. In some embodiments, the metal surface is fully oxidized. In some embodiments, a portion of the metal surface is oxidized.

In some embodiments, at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50% of the metal surface , at least about 60%, at least about 70%, at least about 80%, or at least about 90% are oxidized. In some embodiments, up to about 1%, up to about 2%, up to about 5%, up to about 10%, up to about 20%, up to about 30%, up to about 40%, up to about 50%% of the metal surface , up to about 60%, up to about 70%, up to about 80%, or up to about 90% are oxidized. In some embodiments, about 1%, about 2%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70% of the metal surface, Up to about 80%, about 90%, or substantially all are oxidized. In some embodiments, the degree of oxidation of a metal surface is measured by the percentage of metal atoms on the metal surface that have an oxidation state greater than zero. In some embodiments, the degree of oxidation of a metal surface is measured by the percentage of the number of oxygen atoms that cover the metal surface.

In some embodiments, the metal surface is a metal oxide layer disposed on the outside of the substrate. In some embodiments, the substrate is a metal substrate. In some embodiments, the metal substrate is an unoxidized metal. In some embodiments, the metal substrate is the same metal as the metal oxide layer. In some embodiments, the metal substrate is a non-oxidized version of an oxidized metal layer. In some embodiments, the substrate is a different metal than the metal surface. In some embodiments, the metal oxide layer has a thickness of about 0.1 nm to about 10 nm. In some embodiments, the metal oxide layer is about 0.1 nm to about 0.2 nm, about 0.1 nm to about 0.5 nm, about 0.1 nm to about 1 nm, about 0.1 nm to about 5 nm, about 0 .1 nm to about 10 nm, about 0.2 nm to about 0.5 nm, about 0.2 nm to about 1 nm, about 0.2 nm to about 5 nm, about 0.2 nm to about 10 nm, about 0.5 nm to about 1 nm, about 0 .5 nm to about 5 nm, about 0.5 nm to about 10 nm, about 1 nm to about 5 nm, about 1 nm to about 10 nm, or about 5 nm to about 10 nm. In some embodiments, the metal oxide layer has a thickness of about 0.1 nm, about 0.2 nm, about 0.5 nm, about 1 nm, about 5 nm, or about 10 nm. In some embodiments, the metal oxide layer has a thickness of at least about 0.1 nm, about 0.2 nm, about 0.5 nm, about 1 nm, or about 5 nm. In some embodiments, the metal oxide layer has a thickness of at most about 0.2 nm, about 0.5 nm, about 1 nm, about 5 nm, or about 10 nm. In some embodiments, the metal oxide layer has a thickness of about 0.1 nm to about 1,000 nm. In some embodiments, the metal oxide layer is about 0.1 nm to about 0.5 nm, about 0.1 nm to about 1 nm, about 0.1 nm to about 5 nm, about 0.1 nm to about 10 nm, about 0.1 nm. to about 50 nm, about 0.1 nm to about 100 nm, about 0.1 nm to about 500 nm, about 0.1 nm to about 1,000 nm, about 0.5 nm to about 1 nm, about 0.5 nm to about 5 nm, about 0.5 nm from about 10 nm, from about 0.5 nm to about 50 nm, from about 0.5 nm to about 100 nm, from about 0.5 nm to about 500 nm, from about 0.5 nm to about 1,000 nm, from about 1 nm to about 5 nm, from about 1 nm to about 10 nm, about 1 nm to about 10 nm to about 50 nm, about 1 nm to about 100 nm, about 1 nm to about 500 nm, about 1 nm to about 1,000 nm, about 5 nm to about 10 nm, about 5 nm to about 50 nm, about 5 nm to about 100 nm, about 5 nm to about 500 nm, about 5 nm to about 1,000 nm, about 10 nm to about 50 nm, about 10 nm to about 100 nm, about 10 nm to about 500 nm, about 10 nm to about 1,000 nm, about 50 nm to about 100 nm, about 50 nm to about 500 nm, about It has a thickness of 50 nm to about 1,000 nm, about 100 nm to about 500 nm, about 100 nm to about 1,000 nm, or about 500 nm to about 1,000 nm. In some embodiments, the metal oxide layer has a thickness of about 0.1 nm, about 0.5 nm, about 1 nm, about 5 nm, about 10 nm, about 50 nm, about 100 nm, about 500 nm, or about 1,000 nm. . In some embodiments, the metal oxide layer has a thickness of at least about 0.1 nm, about 0.5 nm, about 1 nm, about 5 nm, about 10 nm, about 50 nm, about 100 nm, or about 500 nm. In some embodiments, the metal oxide layer has a thickness of at most about 0.5 nm, about 1 nm, about 5 nm, about 10 nm, about 50 nm, about 100 nm, about 500 nm, or about 1,000 nm.

In some embodiments, the metal surface comprises a mixture of metals. In some embodiments, the metal surface comprises at least two metals selected from platinum, gold, palladium, titanium, and rhodium, or oxides thereof. In some embodiments, the metal surface comprises at least three metals selected from platinum, gold, palladium, titanium, and rhodium, or oxides thereof.

In some embodiments, the metal surface comprises at least one metal selected from platinum, gold, palladium, titanium, and rhodium, or oxides thereof, and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements.

In some embodiments, the metal surface comprises platinum or its oxide and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements. In some embodiments, the metal surface comprises platinum and the additional material comprises silicon, zirconium, gold, aluminum, titanium, iron, or zinc, or oxides thereof, or any combination thereof. In some embodiments, the metal surface comprises platinum and the additional material comprises silicon or zirconium.

In some embodiments, the metal surface comprises gold or its oxide and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements. In some embodiments, the metal surface comprises gold and the additional material comprises silicon, zirconium, platinum, aluminum, titanium, iron, or zinc, or oxides thereof, or any combination thereof. In some embodiments, the metal surface comprises gold and the additional material comprises silicon or zirconium.

In some embodiments, the metal surface comprises palladium or its oxide and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements. In some embodiments, the metal surface comprises palladium and the additional material comprises silicon, zirconium, gold, platinum, titanium, iron, or zinc, or oxides thereof, or any combination thereof. In some embodiments, the metal surface comprises palladium and the additional material comprises silicon or zirconium.

In some embodiments, the metal surface comprises iridium or its oxide and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements. In some embodiments, the metal surface comprises iridium and the additional material comprises silicon, zirconium, gold, aluminum, platinum, iron, or zinc, or oxides thereof, or any combination thereof. In some embodiments, the metal surface comprises iridium and the additional material comprises silicon or zirconium.

In some embodiments, the metal surface comprises rhodium or its oxide and additional materials. In some embodiments the additional material comprises an additional metal. In some embodiments, the additional material includes additional elements. In some embodiments, the metal surface comprises rhodium and the additional material comprises silicon, zirconium, gold, aluminum, platinum, titanium, or zinc, or oxides thereof, or any combination thereof. In some embodiments, the metal surface comprises rhodium and the additional material comprises silicon or zirconium.

In some embodiments, the metal surface is at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 98% metal by weight. including. In some embodiments, the metal surface comprises about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 98% metal by weight. In some embodiments, the metal surface is up to about 50%, up to about 60%, up to about 70%, up to about 80%, up to about 90%, up to about 95% by weight, or Contains up to about 98% metal.

In some embodiments, the metal surface is at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 98% metal by weight. or its oxide. In some embodiments, the metal surface comprises about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 98% by weight of the metal or any oxide thereof. include. In some embodiments, the metal surface is up to about 50%, up to about 60%, up to about 70%, up to about 80%, up to about 90%, up to about 95% by weight, or containing up to about 98% metal or any oxide thereof.

A metal surface as used herein can be a substantially pure metal or an alloy of different materials. In some embodiments, the metal surface comprises a platinum alloy. Platinum may be alloyed with other metals. In some embodiments, the platinum alloy is iron, nickel, copper, palladium, chromium, iridium, osmium, aluminum, tin, molybdenum, vanadium, niobium, tantalum, zirconium, manganese, ruthenium, gold, or another alloy such as cobalt. contains platinum alloyed with metals of In some embodiments, the platinum alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% platinum . In some embodiments, the platinum alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% platinum .

In some embodiments, the metal surface comprises a gold alloy. Gold may be alloyed with other metals. In some embodiments, gold alloys include nickel, silver, palladium, platinum, rhodium, mercury, copper, zinc, aluminum, cadmium, manganese, gallium, indium, chromium, cobalt, iron, or another metal such as ruthenium. including gold alloyed with In some embodiments, the gold alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% gold. . In some embodiments, the gold alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% gold. .

In some embodiments, the metal surface comprises a palladium alloy. Palladium may be alloyed with other metals. In some embodiments, palladium alloys include palladium alloyed with gold, platinum, iridium, rhodium, ruthenium, osmium, silver, nickel, copper, or manganese. In some embodiments, the palladium alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% palladium. . In some embodiments, the palladium alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% palladium. .

In some embodiments, the metal surface comprises an iridium alloy. Iridium may be alloyed with other metals. In some embodiments, the iridium alloy comprises iridium alloyed with platinum, ruthenium, osmium, nickel, cobalt, copper, titanium, or zirconium. In some embodiments, the iridium alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% iridium. . In some embodiments, the iridium alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% iridium. .

In some embodiments, the metal surface comprises a rhodium alloy. Rhodium may be alloyed with other metals. In some embodiments, rhodium alloys include rhodium alloyed with platinum, molybdenum, iridium, ruthenium, palladium, osmium, titanium, rhenium, gold, or nickel. In some embodiments, the rhodium alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% rhodium. . In some embodiments, the rhodium alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% rhodium. .

In some embodiments, the metal surface comprises an aluminum alloy. Aluminum may be alloyed with other metals. In some embodiments, an aluminum alloy comprises aluminum alloyed with another metal such as copper, magnesium, manganese, silicon, tin, zinc, iron, scandium, or zirconium. In some embodiments, the aluminum alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% aluminum . In some embodiments, the aluminum alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% aluminum .

In some embodiments, the metal surface comprises a titanium alloy. Titanium may be alloyed with other metals. In some embodiments, the titanium alloy is titanium alloyed with another metal such as aluminum, vanadium, tin, niobium, iron, tantalum, zirconium, molybdenum, silicon, manganese, chromium, cobalt, nickel, or copper. including. In some embodiments, the titanium alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% titanium. . In some embodiments, the titanium alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% titanium. .

In some embodiments, the metal surface comprises an iron alloy. Iron may be alloyed with other metals or other elements. In some embodiments, iron is alloyed with another material such as carbon, vanadium, chromium, niobium, titanium, nickel, cobalt, aluminum, manganese, uranium, silicon, or copper. In some embodiments, the iron alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% iron. . In some embodiments, the iron alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% iron. . In some embodiments, the iron alloy is steel.

In some embodiments, the metal surface comprises a zinc alloy. Zinc may be alloyed with other metals. In some embodiments, zinc alloys comprise zinc alloyed with another metal such as copper, tin, nickel, silver, mercury, magnesium, silicon, iron, lead, or aluminum. In some embodiments, the zinc iron alloy comprises at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, or at least 98 wt% zinc. include. In some embodiments, the zinc alloy comprises about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, or about 98 wt% zinc. .

Metal surfaces can be functionalized with any number or combination of surface modifications, reactive groups, or capture moieties provided herein. In some embodiments, the metal surface is functionalized with a single type of surface modifying group, reactive group, or capture moiety. In some embodiments, the metal surface is functionalized with multiple different surface modifying groups, reactive groups, or capture moieties. In some embodiments, the metal surface is modified with multiple copies of the same surface modifying group, reactive group, or capture moiety.

II. Uses of Modified Metal Surfaces Metal surfaces modified as provided herein can be used for any suitable purpose or configured to have any desired properties. Due to the wide range of moieties that can be attached to metal surfaces using the methods and reagents described herein, there is no limit to the uses of such surfaces. Non-limiting examples of potential applications of the modified metal surfaces described herein are provided herein.

Because of their conductive properties, metals such as gold and platinum are frequently used in electrodes, among other uses. Gold and platinum electrodes can be used in electrokinetic assays. However, the presence of electrical currents in these assays can have detrimental electrochemical effects on the sample, including electrolysis reactions that can occur on or near the electrodes, heating, and Including, but not limited to, chaotic fluid motion. In some embodiments, the material can be linked to a metal electrode surface using phosphonate linkages provided herein that can mitigate these effects, and can also be used for cells, bacteria, viruses, It allows performing effective separations of nanoparticles, DNA, and other biomolecules. In some embodiments, a hydrogel is attached to the metal surface of the electrode to prevent deposition of biological material on the electrode surface during an electrokinetic assay. In some embodiments, a hydrogel is attached to the metal surface of the electrode to prevent the biological material from being damaged or degraded during the electrokinetic assay. For this purpose, the hydrogel must have sufficient mechanical strength and be relatively chemically inert so that it can withstand electrochemical effects at the electrode surface without deformation or degradation. In general, hydrogels are sufficiently permeable to small aqueous ions, but keep biomolecules away from the electrode surface.

In some embodiments, the metal surface is configured to contact biological material. In some embodiments, the metal surface is configured to contact a biological fluid (eg, blood). In some embodiments, the metal surface is modified with phosphonate moieties provided herein to impart antibiotic adhesion properties to the metal surface. In some embodiments, a metal surface configured to contact biological material includes a biofouling anti-coating such as a hydrogel, hydrophilic polymer, or hydrophobic polymer.

The metal surfaces modified herein can be configured to be placed within the body of an animal. In some embodiments, the metal surface is configured to be placed within the body of a mammal. In some embodiments, the metal surface is configured to be placed within a human body. In some embodiments, the metal surface is part of a medical implant. In some embodiments, the metal surface is part of a surgical or dental implant. In some embodiments, the metal surface is a surgical or dental implant. Such implants may be used to prevent corrosion of the implant, prevent biofouling on the implant, prevent the growth of bacteria or other cells on the implant, or any other desired surface of the implant. The phosphonates provided herein can be modified to alter properties.

In some embodiments, the metal surface comprises materials linked via phosphonate moieties to prevent cell adhesion to the metal surface. In some embodiments, the material is configured to prevent adherence of bacterial cells to metal surfaces. In some embodiments the material comprises a polymer. In some embodiments, the polymer is grafted onto the metal surface. In some embodiments, the polymer comprises a quaternary ammonium ion-containing polymer (eg, poly 2-dimethylaminoethyl methacrylate). In some embodiments, the polymer is poly 2-dimethylaminoethyl methacrylate. In some embodiments, the polymer is poly 4-vinylpyridine. In some embodiments, the polymer is poly(3-(trimethoxysilyl)propyl methacrylate). In some embodiments, the polymer is a fluorocarbon, a perfluorocarbon polymer. In some embodiments, the material comprises antimicrobial peptides. In some embodiments, the material comprises a biocide chemically bound to the phosphonate. In some embodiments, phosphonate-linked materials are used in combination with additional antimicrobial treatments (eg, surface treatments with preservatives such as chlorhexidine and/or chloroxylenol).

In some embodiments, the metal surface comprises materials linked via phosphonate moieties to prevent marine biofouling. In some embodiments, the material comprises a hydrophobic polymer. In some embodiments, the material comprises a silicone elastomer (eg, polydimethylsiloxane). In some embodiments, the material comprises a mixture of silicone polymers. In some embodiments, the material comprises a hydrophobic polymer. In some embodiments, the hydrophobic polymer is polyethylene, polypropylene, polystyrene, polyvinyl halide, polytetrafluoroethylene, poly(methylmethacrylate), polycarbonate, polyetherurethane, polydimethylsiloxane, or any combination or derivative thereof. including. In some embodiments, the material includes a fouling release coating.

In some embodiments, the metal phosphonate linkages provided herein can be used to immobilize drug moieties on delivery devices having metal surfaces. In some embodiments, the delivery device is a nanoparticle, microparticle, bead, or similar piece with a metal surface. In one aspect, provided herein is a drug delivery device comprising a drug moiety covalently attached to the surface of the device via a phosphonate residue, the surface being platinum or gold. In some embodiments the surface is platinum. In some embodiments the surface is gold. In some embodiments, the surface is palladium, iridium, or rhodium. In some embodiments, the drug moiety is linked to the phosphonate residue via a linker. In some embodiments, the linker is a cleavable linker. In some embodiments, the linker is configured to release the drug moiety. A linker can be configured to release the drug moiety near a desired target, such as a tumor. In some embodiments, the drug delivery device further comprises a targeting moiety such as an antibody or antibody fragment, peptide, or nucleic acid.

In some embodiments, the metal phosphonates provided herein can be used to prepare metal surfaces useful for purification of biological media. Phosphonate linkages to metal surfaces can be used to link multiple binding agents to the surface. In some embodiments, the binding agent can bind the material of interest to the surface. In some embodiments, the biological medium is washed onto the surface and the substance of interest is bound to the surface. The biological medium can then be removed from the surface. In some embodiments, the metal surface is then washed to remove non-specifically bound substances from the surface. The material of interest can then be released from the metal surface and recovered (e.g., by cleaving the linker between the binding agent and the surface, or by altering the properties of the binding agent, e.g., by modifying ). In some embodiments, the binding agent can be configured to bind the substance of non-interest in order to remove it from the biological medium. In some embodiments, the binding agent can be placed on a metal surface that can be easily removed from the medium, such as beads or microparticles. Such particles can be collected by various methods (eg, magnetic field or centrifugation). Collection of particles with undesired material binding can clarify complex biological samples, leaving only the material of interest.

In some embodiments, the metal phosphonate linkages provided herein can be used in the preparation of microarrays. Microarrays can be used for any purpose. In some embodiments, microarrays can be used for DNA sequencing, protein expression analysis, protein-protein interaction screening, or any other biological assay that can be performed in array format. In some embodiments, the array comprises a plurality of features attached to the metal surface via phosphonate residues. In some embodiments, these features include probe moieties. In some embodiments, each feature includes a unique probe portion. In some embodiments, probe moieties comprise nucleic acids, peptides, proteins, antibodies, small molecules, glycans, or any combination thereof. In some embodiments, the probe portion comprises nucleic acids. In some embodiments, nucleic acids comprise DNA or RNA. In some embodiments, the nucleic acid is single stranded. In some embodiments, nucleic acids include aptamers. In some embodiments, the microarray is configured for DNA sequencing. In some embodiments, microarrays are configured for expression analysis. In some embodiments, the expression analysis is protein expression analysis. In some embodiments, the probe portion is configured to capture mRNA. In some embodiments, probe moieties comprise peptides or proteins. In some embodiments, the microarray is configured for protein-protein interaction screening. A microarray can have any number of features. In some embodiments, the microarray comprises at least 10, at least 100, at least 1000, at least 10000, or at least 100000 unique probe features.

III. Methods of Altering Metal Surfaces Also provided herein are methods of producing the modified metal surfaces described herein. In some embodiments, metal surfaces can be modified in a robust and reproducible manner using simple procedures. The simplicity of the methods provided herein allows metal surfaces to be modified with a high degree of control and can be used to further functionalize metal surfaces in a reliable and consistent manner from batch to batch.

In one aspect, provided herein is a method of functionalizing a metal surface. In some embodiments, the method includes depositing a phosphonic acid or phosphonate ester reagent onto the metal surface. In some embodiments, the method further comprises heating the metal surface to bind the phosphonic acid or phosphonate ester reagent with the metal surface. In some embodiments, the metal surface is a platinum surface or its oxide. In some embodiments, the metal surface is a gold surface or its oxide. In some embodiments, the metal surface is palladium, titanium, or rhodium, or oxides thereof. In some embodiments, the metal surface is oxidized.

In some embodiments, the phosphonate reagent comprises a phosphonate moiety. In some embodiments, the phosphonate reagent comprises a phosphonate derivative. In some embodiments, the phosphonic acid derivative is a phosphonate ester. In some embodiments, the phosphonate ester is an alkylphosphonate ester. In some embodiments the phosphonate ester is a methyl or ethyl ester. In some embodiments in which the phosphonate reagent is a phosphonate derivative, the method further comprises adding an acid to the reaction mixture. In some embodiments, the acid is an organic acid (eg, acetic acid, formic acid, etc.). In some embodiments, the acid is an inorganic acid (eg, HCl, HBr, _{H2SO4 ,} etc.). In some embodiments the acid is a strong acid. In some embodiments the acid is a weak acid.

In some embodiments, depositing the phosphonate or phosphonate ester reagent comprises contacting the metal surface with a solution comprising the phosphonate reagent. In some embodiments, depositing the phosphonate reagent comprises contacting the platinum surface with a solution comprising the phosphonate reagent and solvent. Any solvent that can dissolve the phosphonic acid reagent can be used. In some embodiments, solvents include organic solvents, aqueous solvents, or any combination or mixture thereof. In some embodiments the solvent is an organic solvent. In some embodiments, the organic solvent is acetic acid, acetone, acetonitrile, benzene, tert-butyl alcohol, tert-butyl methyl ether, carbon tetrachloride, chloroform, cyclohexane, 1,2-dichloroethane, dichloromethane, diethyl ether, diglyme , 1,2-dimethoxyethane, dimethylacetamide, dimethylformamide, dimethylsulfoxide, dioxane, ethanol, ethyl acetate, ethyl methyl ketone, ethylene glycol, hexane, hexamethylphosphoramide, methanol, nitromethane, pentane, 2-proponal, pyridine , tetrahydrofuran, toluene, xylene, or any combination thereof. In some embodiments, organic solvents include ethanol, tetrahydrofuran, or toluene, or any combination thereof. In some embodiments, organic solvents include ethanol, tetrahydrofuran, or toluene. In some embodiments, the organic solvent comprises ethanol. In some embodiments, the organic solvent comprises tetrahydrofuran. In some embodiments, the organic solvent comprises toluene.

The phosphonic acid or phosphonate ester reagent can be present in solution at any concentration. In some embodiments, reagents are present in solution at concentrations up to about 1 μM, 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1 M. In some embodiments, the reagent is present in solution at a concentration of about 1 μM, 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1 M. In some embodiments, the reagent is present in solution at a concentration of at least about 1 μM, 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1 M. In some embodiments, the reagent is present in solution at a concentration of about 1 mM. In some embodiments, the reagent is present in solution at a concentration of about 1 μM, 10 μM, 100 μM, 1 mM, or 10 mM to about 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1 M. , the reagent is present in solution at a concentration of about 1 μM to about 1M.

In some embodiments, depositing the reagent on the metal surface comprises evaporating the solvent from the metal surface. Any suitable method can be used to evaporate the solvent. In some embodiments, the solvent is allowed to evaporate under atmospheric pressure. In some embodiments, a vacuum source is used to evaporate the solvent. In some embodiments, evaporating the solvent comprises heating the metal surface. In some embodiments, the metal surface is heated to a temperature of at least 30°C, at least 40°C, at least 50°C, at least 60°C, at least 70°C, at least 80°C, or at least 90°C. In some embodiments, the metal surface is heated to a temperature of about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, or about 90°C.

In some embodiments, the metal surface is heated to bind the phosphonic acid or phosphonate ester reagent to the metal surface. Any method of heating the surface can be used. In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface is performed in an oven, vacuum oven, or microwave reactor. In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface is performed in an oven. In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface is performed in a vacuum oven. In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface is performed in a microwave reactor.

In some embodiments, heating the metal surface to bind the phosphonic acid or phosphonate ester reagent to the metal surface is at least 30° C., at least 50° C., at least 70° C., at least 80° C., at least 100° C., at least heating the metal surface to a temperature of 120°C, at least 140°C, at least 160°C, or at least 180°C. In some embodiments, heating the metal surface to bind the reagent with the metal surface is about 30°C, about 50°C, about 70°C, about 80°C, about 100°C, about 120°C, about 140°C. C., about 160.degree. C., or about 180.degree. In some embodiments, heating the metal surface to bind the reagent with the metal surface may be up to 50°C, up to 70°C, up to 80°C, up to 100°C, up to 120°C, up to 140°C, up to 160°C. °C, or up to 180°C to heat the metal surface.

In some embodiments, the metal surface is oxidized. In some embodiments, the metal surface is a metal oxide surface. In some embodiments, the metal surface is partially oxidized. In some embodiments, the metal surface is at least partially oxidized. In some embodiments, the metal surface is fully oxidized.

In some embodiments, the method further comprises oxidizing the metal surface. Oxidation of metal surfaces can be accomplished using any method. In some embodiments, the metal surface is oxidized by air oxidation, plasma treatment, ultraviolet ozone oxidation, or chemical oxidation. In some embodiments, the metal surface is oxidized by air oxidation. In some embodiments, the metal surface is oxidized by ultraviolet ozone oxidation. In some embodiments, the metal surface is oxidized by plasma treatment. In some embodiments, the metal surface is oxidized by chemical oxidation.

Phosphonic acid or phosphonate ester reagents can have any structure. In some embodiments, the phosphonic acid or phosphonate ester reagent has the structure
has
wherein each R ^E is independently H or optionally substituted alkyl; and
R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally or optionally substituted heteroaryl.
In some embodiments, each R ^E is independently H or optionally substituted C ₁ -C ₆ alkyl. In some embodiments, each R ^E is independently H or C ₁ -C ₆ alkyl. In some embodiments, each R ^E is independently H, methyl, ethyl, or propyl. In some embodiments, each ^RE is H. In some embodiments, each R ^E is independently methyl, ethyl, or propyl.

The R group of the phosphonic acid or phosphonate ester reagent can be any group. In some embodiments, R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted is optionally substituted alkynyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, R is any of the groups R described in the "Modified Metal Surfaces" section. In some embodiments, the phosphonate reagent is a phosphonate ester containing any of the R groups described in the "Modified Metal Surfaces" section. In some embodiments, the phosphonate ester is an alkylphosphonate ester. In some embodiments, the phosphonate ester is a C ₁ -C ₆ alkyl phosphonate ester. In some embodiments, the phosphonate ester is a methyl, ethyl, or propyl phosphonate ester.

In embodiments where the phosphonate or phosphonate ester reagents include reactive tags or capture moieties, additional groups can be added to further modify the properties of the metal surface. In some embodiments, the method further comprises adding a second group to the reagent by performing an additional reaction. In some embodiments, additional groups are added after conjugation of the phosphonic acid or phosphonate ester reagent to the metal surface. Any suitable reaction conditions that allow reaction of the reactive tag or capture moiety can be used. In some embodiments, additional groups are added by CLICK chemistry. In some embodiments, additional groups are added by nucleophilic addition. In some embodiments, additional groups are added by polymerization reactions. In some embodiments, the polymerization reaction is (ATRP)-mediated atom transfer radical polymerization. In some embodiments, the polymerization reaction is reversible addition-fragmentation chain transfer (RAFT) polymerization. In some embodiments, additional groups are added by cycloaddition reactions. In some embodiments, additional groups are added by conjugation reactions. In some embodiments, additional groups are added by bioconjugation reactions. The reactions used to add additional groups can be used in any solvent or solution, such as organic solvents, aqueous solutions, or suitable buffers.

IV. DEFINITIONS Unless otherwise defined, all technical terms, notations, and other technical and scientific terms or terminology used herein are commonly understood by those of ordinary skill in the art to which the claimed subject matter pertains. intended to have the same meaning as understood by In some instances, terms with commonly understood meanings are defined herein for clarity and/or ease of reference, and the inclusion of such definitions herein should not necessarily be construed to represent a material difference from that generally understood in the art.

Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. Reference to a range, eg, 1 to 6, refers to subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values within the ranges, eg, 1, 2, 3, 4, 5, and 6 should be considered as specifically disclosing. This applies regardless of the width of the range.

As used in the specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "sample" includes multiple samples, including mixtures thereof.

The terms "determine," "measure," "evaluate," "calculate," "assay," and "analyze" are used interchangeably herein to refer to forms of measurement. It is often done. These terms include determining whether an element is present (eg, detecting). These terms can include quantitative, qualitative, or quantitative and qualitative determinations. Evaluation can be relative or absolute. "Detecting the presence of" can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.

The terms "subject," "individual," or "patient" are often used interchangeably herein. A "subject" can be a biological entity that contains expressed genetic material. A biological entity can be, for example, a plant, animal, or microorganism, including bacteria, viruses, fungi, and protozoa. The subject can be biological entities obtained in vivo or cultured in vitro, tissues, cells and their progeny. A subject can be a mammal. A mammal can be a human. The subject may have been diagnosed or suspected to be at high risk for the disease. In some cases, the subject is not necessarily diagnosed or suspected to be at high risk for the disease.

As used herein, the term "about" a number refers to that number plus or minus 10% of that number. The term "about" a range refers to the range minus the lowest value of 10% plus the highest value of 10%.

"Alkyl" refers to a straight or branched hydrocarbon chain radical consisting exclusively of carbon and hydrogen atoms, which is optionally unsaturated with one or more double or triple bonds. Alkyl is attached to the rest of the molecule through a single bond. Unless otherwise specified, the term "alkyl" and its equivalents include straight or branched chain alkyl groups. When an alkyl group is described as "linear," the mentioned alkyl group is not substituted with additional alkyl groups and is not branched. When an alkyl group is described as "saturated," the alkyl group being referred to does not contain double or triple carbon-carbon bonds (eg, alkenes or alkynes). Alkyl groups include, for example, 1 to 15 carbon atoms (ie C ₁ -C ₁₅ alkyl), 1 to 13 carbon atoms (ie C ₁ -C ₁₃ alkyl), 1 to 8 carbon atoms ( C ₁ -C ₈ alkyl), 1 to 5 carbon atoms (ie C ₁ -C ₅ alkyl), 1 to 3 carbon atoms (ie C ₁ -C ₃ alkyl), or 1 A carbon atom (i.e. C1 _alkyl ). Additional lengths are possible, such as up to 20 carbon atoms, up to 30 carbon atoms, up to 50 carbon atoms, or up to 100 carbon atoms. In certain embodiments, alkyl groups are methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), is selected from 2-methylpropyl, 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl) (isobutyl);

"Alkenyl" is a straight or branched hydrocarbon chain radical group consisting exclusively of carbon and hydrogen atoms and containing at least one carbon-carbon double bond and preferably having from 2 to 12 carbon atoms. (ie C ₂ -C ₁₂ alkenyl)). In certain embodiments, alkenyl contains 2-10 carbon atoms (ie, C ₂ -C ₁₀ alkenyl). In certain embodiments, alkenyl contains 2-8 carbon atoms (ie, C ₂ -C ₈ alkenyl). In other embodiments, the alkenyl contains 2-6 carbon atoms (ie, C ₂ -C ₆ alkenyl). Alkenyl can be attached to the remainder of the molecule by a single bond, for example, ethenyl (ie, vinyl), propenyl (ie, allyl), but-1-enyl, pent-1-enyl, penta-1,4- dienyl and the like.

"Alkynyl" means a straight or branched hydrocarbon chain radical group consisting only of carbon and hydrogen atoms and containing at least one carbon-carbon triple bond and preferably having from 2 to 12 carbon atoms. (ie C ₂ -C ₁₂ alkynyl). ). In certain embodiments, alkynyl contains 2-8 carbon atoms (ie, C ₂ -C ₈ alkynyl). In other embodiments, the alkynyl contains 2-6 carbon atoms (ie, C ₂ -C ₆ alkynyl). In other embodiments, alkynyl contains 2-4 carbon atoms (ie, C ₂ -C ₄ alkynyl). Alkynyl can be attached to the rest of the molecule by a single bond, eg, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

"Alkylene", "alkylene chain", or "alkyl chain" refers to a straight or branched divalent hydrocarbon chain connecting the remainder of the molecule to a radical group consisting only of carbon and hydrogen. , optionally one or more double or triple carbon-carbon bonds, preferably those having 1 to 12 carbon atoms, such as methylene, ethylene, propylene, n-butylene, etc. An alkylene chain is an alkylene chain molecule and the points of attachment to the radical group can be via any two carbons in the chain. In certain embodiments containing 10 carbon atoms (ie, C ₁ -C ₈ alkylene), the alkylene contains 1-8 carbon atoms (ie, C ₁ -C ₈ alkylene). In other embodiments, the alkylene contains 1-5 carbon atoms (ie, C ₁ -C ₈ alkylene). C ₅ alkylene) Other emboalkylenes contain from 1 to 4 carbon atoms (ie, C ₁ -C ₄ alkylene). In other embodiments, an alkylene contains 1-3 carbon atoms (ie, C ₁ -C ₃ alkylene). In other embodiments, an alkylene contains 1-2 carbon atoms (ie, C ₁ -C ₂ alkylene). In other embodiments, an alkylene contains 1 carbon atom (ie, C ₁ alkylene). In other embodiments, the alkylene contains 5-8 carbon atoms (ie, C ₅ -C ₈ alkylene). In other embodiments, the alkylene contains 2-5 carbon atoms (ie, C ₂ -C ₅ alkylene). In other embodiments, the alkylene contains 3-5 carbon atoms (ie, C ₃ -C ₅ alkylene).

An "alkenylene" or "alkenylene chain" consists of carbon and hydrogen for the remainder of the molecule, contains at least one carbon-carbon double bond, and preferably has 2 to 12 carbon atoms. The alkenylene chain is attached to the rest of the molecule through a single bond and to a radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and radical groups can be through any two carbons in the chain. In certain embodiments, the alkenylene contains 2-10 carbon atoms (ie, C ₂ -C ₁₀ alkenylene). In certain embodiments, alkenylene contains 2-8 carbon atoms (ie, C2-C8 alkenylene). In other embodiments, the alkenylene contains 2-5 carbon atoms (ie, C ₂ -C ₅ alkenylene). In other embodiments, the alkenylene contains 2-4 carbon atoms (ie, C ₂ -C ₄ alkenylene). In other embodiments, the alkenylene contains 2-3 carbon atoms (ie, C ₂ -C ₃ alkenylene). In other embodiments, an alkenylene contains 2 carbon atoms (ie, a _C2 alkenylene). In other embodiments, the alkenylene contains 5-8 carbon atoms (ie, C ₅ -C ₈ alkenylene). In other embodiments, the alkenylene contains 3-5 carbon atoms (ie, C ₃ -C ₅ alkenylene).

An "alkynylene" or "alkynylene chain" links the remainder of the molecule to a radical group, consists solely of carbon and hydrogen, contains at least one carbon-carbon triple bond, preferably a divalent hydrocarbon chain. Point. It has 2 to 12 carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to a radical group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to radical groups can be through any two carbons in the chain. In certain embodiments, the alkynylene contains 2-10 carbon atoms (ie, C ₂ -C ₁₀ alkynylene). In certain embodiments, the alkynylene contains 2-8 carbon atoms (ie, C ₂ -C ₈ alkynylene). In other embodiments, the alkynylene contains 2-5 carbon atoms (ie, C ₂ -C ₅ alkynylene). In other embodiments, the alkynylene contains 2-4 carbon atoms (ie, C ₂ -C ₄ alkynylene). In other embodiments, the alkynylene contains 2-3 carbon atoms (ie, C ₂ -C ₃ alkynylene). In other embodiments, an alkynylene contains 2 carbon atoms (ie, a _C2 alkynylene). In other embodiments, the alkynylene contains 5-8 carbon atoms (ie, C ₅ -C ₈ alkynylene). In other embodiments, the alkynylene contains 3-5 carbon atoms (ie, C ₃ -C ₅ alkynylene).

"Aryl" refers to an aromatic monocyclic or polycyclic hydrocarbon ring system. An aromatic monocyclic or aromatic polycyclic hydrocarbon ring system contains only hydrogen and carbon and from 5 to 18 carbon atoms, wherein at least one of the rings of the ring system is aromatic, i.e. , which contains a cyclic delocalized (4n+2) π-electron system according to Hückel theory. Ring systems from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene.

“Aralkyl,” “arylalkyl,” or “arylalkylene” refer to groups of formula R ^c -aryl, where R ^c is an alkylene chain as defined above, eg, methylene, ethylene, and the like. The alkylene chain portion of the aralkyl group may be substituted as described above for alkylene chains.

"Aralkenyl" refers to a group of formula -R ^d -aryl, where R ^d is an alkenylene chain as defined above. The aryl portion of the aralkenyl group is optionally substituted as described above for aryl groups.

"Aralkynyl" refers to a group of formula R ^e -aryl, where R ^e is an alkynylene chain as defined above. The aryl portion of the aralkynyl group may be optionally substituted as described above for aryl groups.

The terms "C _xy " or "C _x -C _y " when used in conjunction with chemical moieties such as alkyl, alkenyl, or alkynyl refer to groups containing x to y carbons in the chain. means to contain For example, the term "C _xy alkyl" refers to saturated or unsaturated hydrocarbon groups, including straight-chain and branched-chain alkyl groups containing x to y carbons in the chain. The terms "C _xy alkenyl" and "C _xy alkynyl" refer to unsaturated alkyl groups analogous in length and possible substitution to the alkyls described above, but containing at least one double or triple bond, respectively. Refers to an aliphatic group.

"Cycloalkyl" refers to a saturated ring in which each atom of the ring is carbon. Cycloalkyls can include monocyclic and polycyclic rings, such as 3-10 membered monocyclic rings, 6-12 membered fused bicyclic rings, 6-12 membered spirocyclic rings, and 6-12 membered bridged rings. In certain embodiments, cycloalkyl contains 3-10 carbon atoms. In other embodiments, the cycloalkyl contains 5-7 carbon atoms. A cycloalkyl can be attached to the rest of the molecule through a single bond. Examples of monocyclic cycloalkyls include, eg, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl (ie, bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7 dimethylbicyclo[2.2.1]heptanyl, and the like. Unless otherwise specified (for example, when "cycloalkyl" is described as "saturated"), the cycloalkyl groups described herein may have one or more carbon-carbon double or triple bonds. can contain.

"Heteroalkyl" refers to straight or branched hydrocarbon chain radicals containing one or more heteroatoms. Heteroalkyl groups can optionally be unsaturated at one or more double or triple bonds between adjacent carbon atoms, between a carbon atom and a heteroatom, or between two heteroatoms. A heteroalkyl is attached to the rest of the molecule through a single bond. Unless otherwise specified, the term "heteroalkyl" and its equivalents include straight or branched chain alkyl groups. When a heteroalkyl group is described as "linear," the referenced heteroalkyl group is not substituted with additional alkyl or heteroalkyl groups and is not branched. When a heteroalkyl group is described as "saturated", the alkyl group to which it is referred does not contain double or triple bonds (eg, alkenes or alkynes). Heteroalkyl groups can have, for example, 1 to 15 atoms, 1 to 13 atoms, 1 to 8 atoms, 1 to 5 atoms, 1 to 3 atoms, or 1 to 2 atoms. can be of any length, including Longer lengths are possible, such as up to 20 atoms, up to 30 atoms, up to 50 atoms, or up to 100 atoms.

A “heteroalkylene,” “heteroalkylene chain,” or “heteroalkyl chain” is unsaturated at carbon, hydrogen, and optionally one or more double or triple carbon-carbon or carbon-heteroatom bonds. I'm here. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. A heteroalkylene chain is attached to the rest of the molecule through a single bond and to a radical group through a single bond. The points of attachment of a heteroalkylene chain to the rest of the molecule and to radical groups can be through any two atoms within the chain.

"Heterocycle" refers to a saturated, unsaturated or aromatic ring containing carbon atoms and one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles may be monocyclic or polycyclic and include 3-10 membered monocyclic rings, 6-12 membered fused bicyclic rings, 6-12 membered spirocyclic rings, and 6-12 membered bridged rings. Each ring of a bicyclic heterocycle may be selected from saturated, unsaturated and aromatic rings. In some embodiments, a heterocycle is a heteroaryl. In some embodiments, a heterocycle is a heterocycloalkyl. In an exemplary embodiment, a heterocyclic ring such as pyridyl can be fused to a saturated or unsaturated ring such as cyclohexane, cyclopentane, or cyclohexene.

"Heterocycloalkyl" refers to a saturated ring having carbon atoms and at least one heteroatom. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycloalkyl can include monocyclic and polycyclic rings, such as 3-10 membered monocyclic rings, 6-12 membered fused bicyclic rings, 6-12 membered spirocyclic rings, and 6-12 membered bridged rings. A heteroatom in the heterocycloalkyl radical is optionally oxidized. If one or more nitrogen atoms are present, it is optionally quaternized. A heterocycloalkyl is attached to the remainder of the molecule through any atom of the heterocycloalkyl, such as any carbon or nitrogen atom of the heterocycloalkyl, valency permitting. Examples of heterocycloalkyl radicals include dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroiso indolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl , thiamorpholinyl, 1-oxothiomorpholinyl, and 1,1-dioxothiomorpholinyl.

"Heterocycloalkenyl" refers to a saturated ring having carbon atoms and at least one heteroatom, with at least one double bond between the two ring carbons. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycloalkenyl includes monocyclic and It may contain polycyclic rings. In other embodiments, the heterocycloalkenyl contains 5-7 ring atoms. A heterocycloalkenyl can be attached to the rest of the molecule through a single bond. Examples of monocyclic cycloalkenyls include, for example, pyrroline (dihydropyrrole), pyrazoline (dihydropyrazole), imidazoline (dihydroimidazole), triazoline (dihydrotriazole), dihydrofuran, dihydrothiophene, oxazoline (dihydrooxazole), isoxazoline (dihydroisoxazole), thiazoline (dihydrothiazole), isothiazoline (dihydroisothiazole)), oxadiazoline (dihydrooxadiazole), thiadiazoline (dihydrothiadiazole), dihydropyridine, tetrahydropyridine, dihydropyridazine, tetrahydropyridazine, dihydropyrimidine , tetrahydropyrimidine, dihydropyrazine, tetrahydropyrazine, pyran, dihydropyran, thiopyran, dihydrothiopyran, dioxin, dihydrodioxin, oxazine, dihydrooxazine, thiazine, and dihydrothiazine.

"Heteroaryl" refers to an aromatic ring containing carbon atoms and one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. As used herein, heteroaryl rings may be selected from monocyclic or bicyclic and fused or bridged ring systems wherein at least one of the rings of the ring system is aromatic. , that is, it contains a cyclic delocalized (4n+2) π-electron system according to the Hückel theory. Heteroatoms in heteroaryl radicals may be optionally oxidized. If one or more nitrogen atoms are present, it is optionally quaternized. A heteroaryl can be attached to the rest of the molecule through any atom of the heteroaryl, such as a carbon or nitrogen atom of the heteroaryl, valency permitting. Examples of heteroaryl include azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][ 1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d ] pyrimidinyl, 6,7-dihydro-5H cyclopenta[4,5]thieno[2,3d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H -benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2c]pyridinyl, 5,6,7,8,9,10 hexahydro Cycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl , imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7 , 8,9-tetrahydro 5H cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8 tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, Examples include, but are not limited to, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (ie, thienyl).

The term "substituted" refers to moieties that have substituents replacing hydrogens on one or more carbon or heteroatoms of the structure. "Substituted" or "substituted with" includes the implied proviso that such substitution is subject to the permissible valences of the substituting atoms and substituent groups, and that the substitution results in a stable compound, e.g., voluntarily As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms. When a particular group is described as "optionally substituted," it is intended that the group may or may not contain substituents. In embodiments where it is not specified whether a group is substituted or unsubstituted, it is contemplated that the group is optionally substituted. Unless otherwise specified, the substituents described herein may be further substituted with their own substituents, each of which is optionally further substituted.

Substituents can include any of the substituents described herein, such as halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, etc.). or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, aralkyl, carbocycle, heterocycle , cycloalkyl, heterocycloalkyl, aromatic and heteroaromatic moieties. In some embodiments, substituents can include any substituents described herein, such as halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN) , nitro (—NO ₂ ), imino (=N—H), oximo (=N—OH), hydrazino (=N—NH ₂ ), —R ^b OR ^a , —R ^b OC(O)R ^a , — R ^b OC(O)OR ^a , —R ^b OC(O)N(R ^a ) ₂ , —R ^b N(R ^a ) ₂ , —R ^b C(O)R ^a , —R ^b C(O) OR ^a , —R ^b C(O)N(R ^a ) ₂ , —R ^b OR ^c C(O)N(R ^a ) ₂ , —R ^b N(R ^a )C(O)OR ^a , —R ^bN (R ^a )C(O)R ^a , R ^b N(R ^a )S(O) _t R ^a (t is 1 or 2), R ^b S(O) _t R ^a (t is 1 or 2 ), R ^b S(O) _t OR ^a (t is 1 or 2), and R ^b S(O) _t N(R ^a ) ₂ (t is 1 or 2), and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl, any of which is alkyl, alkenyl, alkynyl, halogen, hydroxy, haloalkyl, Haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO ₂ ), imino (=N-H), oximo (=N-OH), hydrazine (= N—NH ₂ ), —R ^b OR ^a , —R ^b OC(O)R ^a , —R ^b OC(O) OR ^a , —R ^b OC(O)N(R ^a ) ₂ , —R ^b N (R ^a ) ₂ , —R ^b C(O)R ^a , —R ^b C(O))OR ^a , —R ^b C(O)N(R ^a ) ₂ , —R ^b OR ^c C(O) N(R ^a ) ₂ , —R ^b N(R ^a )C(O)OR ^a , —R ^b N(R ^a )C(O)R ^a , —R ^b N(R ^a )S(O) _t R ^a (t is 1 or 2), R ^b S(O) _t R ^a (t is 1 or 2), R ^b S(O) _t OR ^a (t is 1 or 2) and R ^b S(O) optionally substituted by _tN (R ^a ) ₂ (t is 1 or 2), where each R ^a is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, is independently selected from heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, and each R ^a is alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O ), thioxo (=S), cyano (--CN), nitro (--NO ₂ ), imino (=N--H), oximo (=N--OH), hydrazine (=N--NH ₂ ), R ^b OR ^a , R ^b OC(O)R ^a , R ^b OC(O)OR ^a , R ^b OC(O)N(R ^a ) ₂ , R ^b N(R ^a ) ₂ , R ^b C(O)R ^a , R ^b C(O)OR ^a , R ^b C(O)N(R ^a ) ₂ , R ^b OR ^c C(O)N(R ^a ) ₂ , R ^b N(R ^a )C(O)OR ^a , ^RbN ( ^Ra )C(O)Ra, RbN(Ra)S(O)tRa (t is 1 or 2), ^RbS ( ^O ) _tRa (t is 1 or 2), ^Rb optionally substituted with S(O) _t OR ^a (t is 1 or 2), R ^b S(O) _t N(R ^a ) ₂ (t is 1 or 2), and each R ^b is Independently selected from a direct bond or a straight or branched alkylene, alkenylene, or alkynylene chain, each R ^c is a straight or branched alkylene, alkenylene, or alkynylene chain.

Unless otherwise stated, it is intended that any group with multiple points of attachment to a parent molecule or two or more moieties can be oriented in any direction. For example, if B
For structures of formula ABC where
are intended to be encompassed.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

V. EXAMPLES The following examples are included for illustration only and are not intended to limit the scope of the invention.

Example 1: Functionalization of a platinum surface with (11-((2-bromo-2-methylpropanoyl)oxy)undecyl)phosphonic acid To functionalize a platinum surface in order to bind phosphonic acid reagents to the platinum surface various conditions were tried. The reagent (11-((2-bromo-2-methylpropanoyl)oxy)undecyl)phosphone (BMPOUA) was chosen because it contains a 2-bromo-2-methylpropanoyl moiety, which This was because the platinum surface could be further functionalized after direct attachment of the phosphonic acid moiety of .

Initial experiments utilized the protocol shown in FIG. Briefly, 40 mg of BMPOUA was dissolved in 100 mL of tetrahydrofuran (THF). A platinum tip was then added to the THF solution. The THF solution was then heated to 60° C. to evaporate the THF. The chips were placed in an oven at 140° C. for 2 days to anneal the phosphonic acid to the surface. The chip was then rinsed with methanol and dried.

To evaluate the effectiveness of the above reaction conditions for attaching phosphonic acid reagents to surfaces, the 2-bromo-2-methylpropanoyl moiety of the resulting metal phosphonate was converted to a suitable acrylate reagent in a radical polymerization reaction. I tried to connect with If the platinum surface was successfully modified with BMPOUA, the polymer should "grow" on the platinum surface under reaction conditions. Polymer growth was observed on the chips, but with considerable variability from chip to chip.
Next, the influence of the solvent on the reaction was investigated. Solutions of 1 mM phosphonic acid were prepared using THF, ethanol (EtOH), or toluene as solvents. A platinum tip was then immersed in the mixture at 60° C. without mixing. The reaction was run for 22 hours and included evaporating the reaction solvent, removing the platinum chip from the solvent and rinsing with methanol, and removing the platinum from the chip and placing it directly in a vacuum oven at 140°C overnight. , processed in various ways. Following these surface modification protocols, suitable acrylate reagents were tried to grow polymers on the surface of the chip. EtOH was found to be the most robust solvent for surface functionalization reactions as it showed the most consistent amount of polymer growth following the initial surface functionalization.

Example 2 Effect of Pt Surface Oxidation on Functionalization The presence of —OH groups on the platinum surface was suspected to play a role in the covalent binding of phosphonates. To evaluate the role of platinum oxidation state in covalent bond formation with phosphonic acids, platinum chips were subjected to plasma treatment (high-energy _O2 plasma reactive ion etching) to oxidize and etch part of the platinum surface. Either treatment with a post-stripping reagent (eg, EKC4000® from DuPont) was performed to remove oxidized platinum, or the platinum tip was not subjected to any treatment prior to reaction with the phosphonic acid reagent.

Plasma-treated and untreated platinum tips were analyzed by X-ray photoelectron spectroscopy (XPS) to assess the oxidation state of the tips. Atomic composition and chemical state information were obtained with a PHIQuantum 2000 using the following parameters:
X-ray source: Monochromated Alkα 1486.6 eV; acceptance angle: ±23°; take-off angle: 45°; analysis area: 600 μm diameter. Charge correction: C1s 284.8 eV (CC, CH). Assignments of chemical states of specific elements were made by referencing reference data from the literature. Non-linear least squares (NLLS) curve fitting was applied to assist in chemical state assignments.

XPS measurements of oxidized and untreated platinum tips are shown in FIGS. 3A and 3B. FIG. 3A shows a survey scan of the surface of three chips. Chips 1 and 2 were subjected to plasma treatment and chip 3 was untreated. FIG. 3B shows an XPS evaluation of various oxidation states of platinum on the chip. The data show that platinum exists in various oxidation states on each chip. The relative proportions of platinum in each oxidation state (Pt0, Pt ²⁺ , Pt ⁴⁺ ) for oxidized platinum and non-plasma treated tips are shown in Table 1 below. In the untreated chip, most of the platinum remained unoxidized. In contrast, plasma-treated platinum tips showed increased levels of oxidized platinum.

After evaluating the platinum tip for oxidation level, the surface was subjected to a functionalization reaction with BMPOUA. BMPOUA (Millipore Sigma) was dissolved in ethanol (Millipore Sigma) at a concentration of 1.2 mM in a round bottom flask. After mixing for 5 minutes at room temperature, the solution was poured into a reaction vessel and a platinum metal substrate (plasma treated, untreated, or post-etch removal reagent treated) was submerged in the phosphonic acid solution without agitation. The reaction vessel was flushed with ultrapure argon gas (Airgas) for 5 minutes and the platinum substrate was left immersed for 18 hours at room temperature. After completion, the substrate was removed from the ethanol solution and placed directly into an oven (VWR Gravity Convection Oven) to anneal the BMPOUA to the substrate at 140° C. for 2 hours without washing.

To evaluate the amount of BMPOUA annealed to the platinum substrate, the material was subjected to a polymerization reaction with an acrylate reagent. If the platinum surface was successfully modified with BMPOUA, the polymer should "grow" on the platinum surface under reaction conditions. The more BMPOUA bound to the surface of the platinum substrate, the more polymer bound to the tip after reaction. Following the polymerization reaction on the modified surface of plasma-treated, untreated, and post-etch removal reagent treated surfaces, surface roughness measurements were taken at normal scan speeds (backscan=0 mm, scan length= Acquired by an optical profilometer (Profilm 3D, Filmetrics F40 microscope) using 0.02 mm, scan average=4). Images were acquired in PSI mode using a 50x DI (Nikon) with a 4x zoom. Following image acquisition, a spline filter (Gaussian β=0.625242, cutoff length=8 μm) was applied. The resulting average polymer hydrogel thickness measurements for plasma-treated and untreated chips are shown in Table 2 below. The platinum tip treated with the post-etch stripper showed no observable polymer growth. Comparing the coefficients of variation and average thickness, the more oxidized surface showed a more uniform polymer (oxidized PtCV% = 6.1 vs. non-oxidized PtCV% = 11.0) and a thicker polymer (average thickness of oxidized Pt polymer thickness = 33.1 nm vs. average thickness of unoxidized Pt polymer = 26.1 nm). In addition, the ability of functionalized platinum substrates to conduct electrical current was evaluated. As expected, the untreated metal substrates showed higher current measurements (108.4 mAmps) compared to the more oxidized plasma treated substrates (average = 99.2 mAmps).

Example 3 Characterization of Modified Platinum Surfaces Platinum chips were functionalized with BMPOUA and characterized using various techniques. The functionalization protocol is as follows: BMPOUA (Millipore Sigma) was dissolved in ethanol (Millipore Sigma) at a concentration of 1.2 mM in a round bottom flask. After mixing for 5 minutes at room temperature, the solution was poured into a reaction vessel and a platinum metal substrate (plasma treated, untreated, or post-etch removal reagent treated) was submerged in the phosphonic acid solution without agitation. The reaction vessel was flushed with ultrapure argon gas (Airgas) for 5 minutes and the platinum substrate was left immersed for 18 hours at room temperature. After completion, the substrate was removed from the ethanol solution and directly placed in an oven (VWR Gravity Convection Oven) at 140° C. for 2 hours without washing to anneal the BMPOUA to the substrate.

After reaction with BMPOUA, evidence of platinum metal functionalization with phosphonic acid was observed by measuring the arithmetic mean height (Sa) before and after functionalization. This was accomplished using an optical profilometer (Profilm3D, Filmetrics). Surface roughness measurements were taken using normal scan speeds (backscan=0 mm, scan length=0.02 mm, scan average=4). Images were acquired in PSI mode using a 50x DI (Nikon) with a 4x zoom. Following image acquisition, images were cropped such that only the platinum electrode was considered and a spline filter (Gaussian β=0.625242, cutoff length=8 μm) was applied. As shown in Table 3, treatment of platinum metal with phosphonic acid increases the surface roughness value by 70% (Sa = 0.94 nm vs. 1.71 nm, bare and treated Pt metal, respectively). . This roughness change can be attributed to the covalent binding of the phosphonic acid to the platinum metal.

FIG. 4 shows an image of a bare platinum electrode prior to surface functionalization. In the platinum electrode of FIG. 4, the arithmetic mean height (Sa) before functionalization is 0.763 nm. FIG. 5 shows the same platinum electrode after functionalization with the BMPOUA phosphonate reagent as described above. The Sa electrode increased to 2.328 nm after functionalization, consistent with successful reagent deposition.

After successful attachment of the phosphonic acid derivative BMPOUA, the functionalized platinum electrode surface was subjected to radical polymerization conditions in the presence of a suitable acrylate reagent. Once the reaction was complete, the surface of the platinum electrode was analyzed by scanning electron microscopy (SEM) as was the platinum electrode that had not undergone derivatization. Images were acquired using an FEI Quanta 250 SEM with a standard working distance of 10 mm. Usually, the acceleration voltage was set to 3-5 keV and the beam current was set to 0.1 nA. All images were obtained using a standard electronic detector (ETD) and the stage was tilted at various angles as shown in Figures 6A, 6B, and 7-14.

FIG. 6A shows an SEM image of a bare platinum electrode surrounded by dielectric at a tilt angle of 45°. Both the platinum metal surface and the dielectric appear smooth. FIG. 6B shows, for comparison, a platinum electrode functionalized with BMPOUA followed by a polymerization reaction similarly surrounded by a dielectric material. After passing a current through the electrodes in FIG. 6B, polymer "folding" is observed at the edge of the modified platinum surface near the interface of the dielectric material.

Figures 7-10 show additional SEM images of polymer-functionalized platinum electrodes. FIG. 7 shows an SEM image taken from a tilt angle of 0°. A folded polymer can be seen on the platinum surface. FIG. 8 shows an SEM image taken at a tilt angle of 14°. The folding of the polymer near the interface of the platinum surface and the dielectric layer can be clearly seen. FIG. 9 shows a similar SEM image at a tilt angle of 14°. The captured area was the same as in Figure 8, but with a wider field of view. FIG. 10 shows an additional SEM image at a tilt angle of 14° from a more zoomed out view.

Figures 11-14 show SEM images of unfunctionalized platinum electrodes surrounded by a dielectric material. Notably, all images show a smooth surface compared to the functionalized platinum surface shown in Figures 7-10. FIG. 11 shows an SEM image at a tilt angle of 1°. The left side of the image shows the platinum surface with the dielectric on the right side. Figure 12 shows an SEM image of the same electrode at a tilt angle of 45°. The interface between the platinum surface and the dielectric material appears smooth. FIG. 13 shows an SEM image at a 4° tilt angle in a zoomed out view of the electrodes. Figure 14 shows a close-up image of the smooth platinum surface interface with the dielectric material at an angle of 45°.

While preferred embodiments of the present disclosure have been shown and described herein, it should be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, modifications, and substitutions will occur to those skilled in the art without departing from this disclosure. It should be understood that various alternatives to the disclosed embodiments described herein may be employed in practicing the disclosure. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims (107)

structure

A functionalized metal surface comprising a metal surface bound to a molecule having
During the ceremony,
R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally aryl substituted with or optionally substituted heteroaryl,
The metal surface is a functionalized metal surface comprising at least one metal selected from platinum, gold, palladium, iridium, and rhodium, oxides thereof, or any combination thereof. R has the structure XL-, where L is a linking moiety and X is a reactive handle or capture moiety, or R has the structure YL-, L is a linking moiety, and Y 2. The functionalized metal surface of claim 1, wherein is a surface-modifying group. 3. The functionalized metal surface of claim 2, wherein L is an optionally substituted alkylene chain or an optionally substituted heteroalkylene chain. L is the formula -[Z-(CR ¹ R ² ] _n ] _m -
has the structure of
wherein each R ¹ and R ² is independently H or optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted or optionally substituted heteroaryl, or R ¹ and R ² on the same atom taken together form oxo, or R ¹ on adjacent atoms taken together forming a double bond or R ¹ and R ² on adjacent atoms together form a triple bond,
each Z is independently absent, -O-, -NR ³ -, -S-, -SS-, -S(O)-, -S(O) ₂ -, -C(O)-, - C(O)O—, —C(O)NR ³ , —OC(O)O—, —NR ³ C(O)NR ³ —, —OC(O)NR ³ —, —S(O) ₂ O -, -S(O) ₂ NR ³ -, -OS(O) ₂ O-, -NR ³ SO ₂ NR ³ -, or -OSO ₂ NR ³ -;
Each R ³ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl,
4. The functionalized metal surface of claim 2 or 3, wherein each n is independently an integer from 1-30, and m is an integer from 1-30. Functionalization according to claim 2 or 3, wherein L comprises a chain of 1-100 atoms, each atom in said chain being independently selected from C, N, O, S, Si, and P. polished metal surface. 6. The functionalized metal surface of claim 5, wherein the first atom in said chain is C. L is
7. The functionalized metal surface of any one of claims 2-6, comprising one or more subunits selected from, wherein each n is independently an integer from 1-30. L is structure
wherein each n is independently an integer from 1 to 30, and each p is independently an integer from 1 to 30 surface. 3. The functionalized metal surface of claim 2, wherein L comprises a polymer. The polymer is polyethylene, polypropylene, poly(vinyl halide), polystyrene, nylon, polyamide, polyester, polyaramid, polyacrylate, polymethacrylate, polytetrafluoroethylene, polysaccharide, poly(alkylene oxide), poly(vinylpyrrolidone) , poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine), or any combination or derivative thereof. A functionalized metal surface according to any one of claims 2 to 10, wherein R has the structure XL-. X is azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, acid, activated ester, alkene, alcohol, halide, acyl halide, sulfonic acid, sulfinic acid, sulfonyl halide, epoxide, aldehyde , ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, and hydrazides, or any combination thereof. X has the structure X ¹ -L ^X -Z ^X -
has
In the formula, Z ^X is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)-, -C( O)O—, —C(O)NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O—, —S(O) ₂ NR ⁴ —, —OS(O) ₂ O—, —NR ⁴ S(O) ₂ NR ⁴ —, or —OS(O) ₂ NR ⁴ —;
L ^X is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted arylene , optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene;
X ¹ is an azide, alkyne, tetrazine, halide, sulfhydryl, disulfide, maleimide, amine, carboxylic acid, active ester, alkene, alcohol, acyl halide, sulfonic acid, sulfinic acid, sulfonyl halide, epoxide, aldehyde, ketone , imines, oximes, isocyanates, isothiocyanates, hydrazines, or hydrazides, and each ^R4 is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl , optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl metal surface. 14. The functionalized compound of claim 13, wherein Z ^X is a bond, -O-, -C(O)-, -NR ⁴ -, -C(O)O-, or -C(O)NR ⁴ metal surface. L ^X is the structure -(CR ⁵ R ⁶ ) _r -
has
wherein each ^R5 and ^R6 is independently H, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally is substituted aryl, or optionally substituted heteroaryl, or R ⁵ and R ⁶ on the same atom taken together form oxo, or R ⁵ taken together on adjacent atoms forming a double bond, or R ⁵ and R ⁶ on adjacent atoms together form a triple bond, and r is an integer from 0 to 30,
15. A functionalized metal surface according to claim 13 or 14. A functionalized metal surface according to any one of claims 13 to 15, wherein X ¹ is an amine, carboxylic acid, maleimide, azide, alkyne, halide, alkene or sulfhydryl. X is
and
During the ceremony,
R ⁵ and R ⁶ are each independently H or C ₁ -C ₃ alkyl;
r is 0 or 1 and X ¹ is an azide, alkyne, halide, alkene, or sulfhydryl;
A functionalized metal surface according to any one of claims 11-16. X is
The functionalized metal surface according to any one of claims 11 to 17, which is 12. The functionalized metal surface of claim 11, wherein X comprises a capture moiety selected from biotin, avidin, streptavidin, nucleic acids, peptides, and proteins, or any combination thereof. A kit comprising a functionalized metal surface according to any one of claims 11-19 and instructions for use. A functionalized metal surface according to any one of claims 2 to 10, wherein R has the structure YL-. Y is a hydrophobic residue, a hydrophilic residue, a charged residue, a cationic residue, an anionic residue, a polysaccharide, a hydrophobic polymer, a hydrophilic polymer, an antibacterial agent, a biological material, a biocompatible material, an antimicrobial 22. The functionalized material of claim 21 comprising surface modifying groups selected from fouling materials, conductive materials, semiconducting materials, heat resistant materials, corrosion inhibiting materials, catalysts, and magnetic materials, or any combination thereof. metal surface. Y has the structure Y ¹ -L ^Y -Z ^Y -
has
In the formula, Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)-, -C(O ) O—, —C(O)NR ⁴ , —OC(O)O—, —NR ⁴ C(O)NR ⁴ —, —OC(O)NR ⁴ —, —S(O) ₂ O—, — S(O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -, or azide, alkyne, tetrazine, halide, sulfhydryl , disulfides, maleimides, amines, carboxylic acids, activated esters, alkenes, alcohols, acyl halides, sulfonic acids, sulfinic acids, sulfonyl halides, epoxides, aldehydes, ketones, imines, oximes, isocyanates, isothiocyanates, hydrazines, or a hydrazide, and a suitable complementary reactive group;
L ^Y is absent, optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted arylene, optionally substituted arylalkylene, optionally substituted heteroarylalkylene, optionally substituted arylheteroalkylene, or optionally substituted heteroarylheteroalkylene;
Y ¹ contains at least one surface modification residue, and each R ⁴ is independently H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted 23. The functionalized metal surface of claim 21 or 22, which is substituted alkynyl, optionally substituted heteroalkyl, optionally substituted aryl, or optionally substituted heteroaryl. Z ^Y is a bond, -O-, -NR ⁴ -, -S-, -SS-, -S(O)-, S(O) ₂ -, -C(O)-, -C(O)O -, -C(O)NR ⁴ -, -OC(O)O-, -NR ⁴ C(O)NR ⁴ -, -OC(O)NR ⁴ -, -S(O) ₂ O-, -S (O) ₂ NR ⁴ -, -OS(O) ₂ O-, -NR ⁴ S(O) ₂ NR ⁴ -, -OS(O) ₂ NR ⁴ -,
24. The functionalized metal surface of claim 23, wherein 25. The functionalized metal surface of claim 23 or 24, wherein ^LY is absent. A functionalized metal surface according to any one of claims 23-25, wherein Y ¹ comprises a hydrophobic residue. 27. The functionalized metal surface of claim 26, wherein said hydrophobic residue comprises a hydrophobic polymer. 4. The hydrophobic polymer comprises polyethylene, polypropylene, polystyrene, polyvinyl halide, polytetrafluoroethylene, poly(methylmethacrylate), polycarbonate, polyether-urethane, polydimethylsiloxane, or any combination or derivative thereof. 27. Functionalized metal surfaces according to 27. A functionalized metal surface according to any one of claims 26-28, wherein said hydrophobic residues comprise fatty acids or derivatives thereof. A functionalized metal surface according to any one of claims 23-25, wherein Y ¹ comprises a hydrophilic residue. 31. The functionalized metal surface of claim 30, wherein said hydrophilic residue comprises a hydrophilic polymer. The hydrophilic polymer is any of polyacrylamide, polyacrylate, polymethacrylate, poly(alkylene oxide), poly(vinylpyrrolidone), poly(vinyl alcohol), polyoxazoline, poly(N-acryloylmorpholine), or derivatives thereof. 32. The functionalized metal surface of claim 31, comprising combinations. The hydrophilic polymer is polyacrylamide, poly(N,N-diethylacrylamide), poly(N,N-dimethylacrylamide), poly(N-isopropylacrylamide), poly(acrylic acid), poly(methacrylic acid), poly (methyl acrylate), poly (ethyl acrylate), poly (2-hydroxyethyl acrylate), poly (propyl acrylate), poly (butyl acrylate), poly (methyl methacrylate), poly (2-hydroxyethyl methacrylate), poly (tetrahydro furfuryl methacrylate), poly(ethylene oxide), poly(propylene oxide), poly(vinylpyrrolidone), poly(oxazoline), poly(2-ethyloxazoline), or poly(N-acryloylmorpholine), claim 31 or 32. A functionalized metal surface according to 32. Y is structure
including
During the ceremony,
each R ⁷ is independently H, optionally substituted C ₁ -C ₃ alkyl, or optionally substituted C ₁ -C ₃ hydroxyalkyl;
each R ⁸ is independently H or C ₁ -C ₃ alkyl;
each X ⁷ is independently absent, -O-, -S-, or -NR ⁹ ;
each R ⁹ is independently H or C ₁ -C ₆ alkyl, and s is an integer from 1 to 10000;
A functionalized metal surface according to any one of claims 21-25. A functionalized metal surface according to any one of claims 23-25, wherein Y ¹ comprises a cationic residue. 36. The functionalized metal surface of claim 35, wherein ^Y1 comprises a cationic polymer. 37. The functionalized metal surface of claim 35 or 36, wherein said cationic residues comprise protonated amine groups, protonated substituted amine groups, quaternary amine groups, or any combination thereof. A functionalized metal surface according to any one of claims 23-25, wherein Y ¹ comprises an anionic residue. 39. The functionalized metal surface of claim 38, wherein ^Y1 comprises an anionic polymer. 40. The functionalized metal surface of claim 38 or 39, wherein said anionic residues comprise carboxylates, sulfonates, sulfinates, phosphates, phosphonates, or any combination thereof. The surface-modifying residue is a hydrophobic residue, a hydrophilic residue, a charged residue, a cationic residue, an anionic residue, a polysaccharide, a hydrophobic polymer, a hydrophilic polymer, an antibacterial agent, a biomaterial, and a biocompatible residue. material, antifouling material, conductive material, semiconducting material, heat resistant material, anti-corrosion material, catalyst, magnetic material, or any combination thereof. Functionalized metal surface. The functionalized metal surface of any one of claims 23-25, wherein said surface modifying groups comprise hydrogels. 43. The functionalized metal surface of claim 42, wherein said hydrogel comprises acrylates, methacrylates, acrylamides, or methacrylamides, or any combination thereof. 44. The functionalized metal surface of claim 42 or 43, wherein said hydrogel has a thickness of about 0.001 microns to about 10 microns. 45. The functionalized metal surface of any one of claims 42-44, wherein said hydrogel has a conductivity of about 0.1 S/m to about 10 S/m. at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least 46. The functionalized metal surface of any one of claims 1-45, wherein about 70%, at least about 80%, or at least about 90% is functionalized. 47. The functionalized metal surface of any one of the preceding claims, wherein said metal surface is at least partially oxidized. at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least 48. The functionalized metal surface of claim 47, wherein about 70%, at least about 80%, or at least about 90% is oxidized. 49. The functionalized metal surface of any one of claims 1-48, wherein said metal surface comprises a metal alloy. 50. The functionalized material of claim 49, wherein the metal alloy comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the metal by weight. metal surface. 51. The functionalized metal surface of any one of claims 1-50, wherein the metal surface comprises a metal and an additional material. 11. The metal surface comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% by weight of a metal or any oxide of a metal. 52. The functionalized metal surface of any one of clauses -51. 53. The functionalized metal surface of any one of claims 1-52, wherein the metal surface is configured to be placed within the body of a mammal. A functionalized metal surface according to any one of the preceding claims, wherein said metal surface is a surgical or dental implant. 55. The functionalized metal surface of any one of claims 1-54, wherein the metal surface is an electrode, microchip, bead, microparticle or nanoparticle. A functionalized metal surface according to any one of the preceding claims, wherein said metal is platinum or gold, or oxides thereof. A functionalized metal surface according to any one of the preceding claims, wherein said metal is palladium, iridium or rhodium, or oxides thereof. A method of functionalizing a metal surface comprising:
(a) depositing a phosphonic acid or phosphonate ester reagent onto a metal surface;
(b) heating the metal surface to bind a phosphonic acid or phosphonate ester reagent to the metal surface;
The method wherein said metal surface comprises platinum, gold, palladium, iridium, or rhodium, oxides thereof, or any combination thereof. 59. The method of claim 58, wherein attaching the phosphonic acid or phosphonate ester reagent comprises contacting the metal surface with a solution comprising a phosphonic acid reagent and a solvent. 60. The method of any one of claims 58 or 59, wherein the solvent comprises an organic solvent, an aqueous solvent, or any combination or mixture thereof. The organic solvent is acetic acid, acetone, acetonitrile, benzene, tert-butyl alcohol, tert-butyl methyl ether, carbon tetrachloride, chloroform, cyclohexane, 1,2-dichloroethane, dichloromethane, diethyl ether, diglyme, 1,2-dimethoxy. Ethane, dimethylacetamide, dimethylformamide, dimethylsulfoxide, dioxane, ethanol, ethyl acetate, ethyl methyl ketone, ethylene glycol, hexanes, hexamethylphosphoramide, methanol, nitromethane, pentanes, 2-proponal, pyridine, tetrahydrofuran, toluene , xylene, or any combination thereof. 61. The method of claim 60, wherein the organic solvent comprises ethanol, tetrahydrofuran, or toluene. 63. The method of any one of claims 59-62, wherein the phosphonate or phosphonate ester reagent is present in solution at a concentration up to about 1 μM, 10 μM, 100 μM, 1 mM, 10 mM, 100 mM, or 1M. 64. The method of any one of claims 59-63, wherein the step of attaching the phosphonic acid or phosphonate ester reagent further comprises evaporating the solvent from the metal surface. 65. The method of Claim 64, wherein evaporating the solvent from the metal surface comprises heating the metal surface. 66. The method of claim 65, wherein the metal surface is heated to a temperature of at least 30°C, at least 40°C, at least 50°C, at least 60°C, at least 70°C, at least 80°C, or at least 90°C. 67. The step of any one of claims 58-66, wherein heating the metal surface to bind the phosphonic acid reagent with the metal surface is performed in an oven, a vacuum oven, or a microwave reactor. Method. heating the metal surface to bind the phosphonic acid or phosphonate ester reagent with the metal surface, heating the metal surface to at least 30° C., at least 50° C., at least 70° C., at least 80° C., at least 100° C., at least 68. The method of any one of claims 58-67, comprising heating to a temperature of 120°C, at least 140°C, at least 160°C, or at least 180°C. A method according to any one of claims 58 to 68, wherein said metal surface is a metal oxide surface. 70. The method of claim 69, wherein the oxidized metal surface is oxidized by air oxidation, plasma treatment, ultraviolet ozone oxidation, or chemical oxidation. 71. The method of any one of claims 58-70, further comprising oxidizing the metal surface. 72. The method of claim 71, wherein oxidizing the metal surface comprises plasma treatment, ultraviolet ozone oxidation, or chemical oxidation. wherein the phosphonic acid or phosphonate ester reagent has the structure
has
During the ceremony,
R is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally aryl substituted with or optionally substituted heteroaryl;
73. The method of any one of claims 58-72, wherein each R ^E is independently H or substituted alkyl. 74. The method of any one of claims 58-73, wherein the metal surface comprises platinum or gold, or oxides thereof. 74. The method of any one of claims 58-73, wherein the metal surface comprises palladium, iridium, or rhodium, or oxides thereof. A metal surface microarray comprising probe moieties attached to a metal surface via phosphonate residues, wherein said metal surface is platinum, gold, palladium, iridium, or rhodium, or oxides thereof, or any combination thereof A metal surface microarray, comprising: The attached phosphonate residue is the structure

wherein R includes the probe moiety and each

77. The microarray of claim 76, wherein , denotes attachment points to the surface. 78. The microarray of claim 76 or 77, wherein said probe moieties comprise nucleic acids, peptides, proteins, antibodies, small molecules, glycans, or any combination thereof. 79. The microarray of claim 78, wherein said probe moieties comprise nucleic acids. 80. The microarray of claim 79, wherein said nucleic acids comprise DNA or RNA. 81. The microarray of any one of claims 76-80, configured for DNA or RNA sequencing. 82. The microarray of any one of claims 76-81, wherein said probe moieties bind to biological agents. 83. The microarray of claim 82, wherein said biological agent is a nucleic acid, protein, cell, or organelle. 84. The microarray of any one of claims 76-83, wherein said probe moieties are specific for biological agents. 85. The microarray of any one of claims 76-84, wherein the microarray comprises at least 10, at least 100, at least 1000, at least 10000, or at least 100000 unique probe features. 86. The microarray of any one of claims 76-85, wherein said metal surface microarray comprises platinum or gold, or oxides thereof. 87. The microarray of any one of claims 76-86, wherein said metal surface microarray comprises palladium, iridium or rhodium, oxides thereof, or any combination thereof. A drug delivery device comprising a drug moiety linked to a surface of the device via a phosphonate residue, said surface comprising platinum, gold, palladium, iridium, or rhodium, oxides thereof, or combinations thereof , drug delivery devices. Phosphonate residues form the structure

wherein R comprises the drug moiety and each

89. A drug delivery device according to claim 88, wherein , denotes attachment points to the surface. 90. The drug delivery device of claim 88 or 89, wherein the drug moiety is linked to the phosphonate residue via a linker. 91. The drug delivery device of claim 90, wherein said linker is a cleavable linker. 92. The drug delivery device of claim 90 or 91, wherein said linker is configured to release said drug moiety. The drug delivery device of any one of claims 88-92, further comprising a targeting moiety. 94. The drug delivery device of claim 93, wherein said targeting moiety comprises an antibody or antibody fragment, peptide, or nucleic acid. 95. The drug delivery device of any one of claims 88-94, which is a nanoparticle, microparticle or bead. 96. The drug delivery device of any one of claims 88-95, wherein the surface comprises platinum or gold, oxides thereof, or combinations thereof. 97. The drug delivery device of any one of claims 88-96, wherein the surface comprises palladium, iridium, or rhodium, oxides thereof, or combinations thereof. structure

A functionalized metal surface comprising a metal surface that binds to a molecule having
During the ceremony,
R is a functionalized metal surface, including hydrogel. platinum, gold, palladium, iridium, rhodium, silicon, aluminum, titanium, iron, zinc, zirconium, nickel, silver, copper, cobalt, or chromium, or oxides thereof, or any combination thereof A functionalized metal surface according to 98. 100. The functionalized metal surface of claim 98 or 99, wherein said hydrogel has a conductivity of about 0.001 S/m to about 10 S/m. 101. The functionalized metal surface of any one of claims 98-100, wherein said hydrogel has a thickness of about 0.001 microns to about 10 microns. 102. The functionalized metal surface of any one of claims 98-101, wherein said hydrogel comprises a synthetic polymer. 103. The functionalized metal surface of any one of claims 98-102, wherein said hydrogel comprises an acrylamide polymer. 104. The functionalized metal surface of any one of claims 98-103, wherein said acrylamide polymer is N-substituted acrylamide, N-substituted methacrylamide, methacrylamide, or any combination thereof. The hydrogel is polyacrylamide, poly(diethylacrylamide), poly(dimethylacrylamide), poly(N-isopropylacrylamide), poly(acrylic acid), poly(methacrylic acid), poly(methyl acrylate), poly(ethyl acrylate) , poly(2-hydroxyethyl acrylate), poly(propyl acrylate), poly(butyl acrylate), poly(methyl methacrylate), poly(2-hydroxyethyl methacrylate), poly(tetrahydrofurfuryl methacrylate), poly(ethylene oxide), 105. The functionalized polymer of any one of claims 98-104, comprising poly(propylene oxide), poly(vinylpyrrolidone), polyoxazoline, poly(2-ethyloxazoline), or poly(N-acryloylmorpholine) metal surface. 106. The functionalized metal surface of any one of claims 98-105, wherein said metal surface is an electrode. 106. The functionalized metal surface of any one of claims 98-105, wherein said metal surface is disposed on the outer surface of an electrode.