Polymerizable compounds integrally containing antibodies and their uses in polymerization induced separation immunoassays

Claim 2

A conjugate used in the de novo preparation of polymers containing one or more polypeptides, comprising: a polymerizable compound containing at least one reactive site for bonding with a polypeptide; and a polypeptide covalently bonded thereto.

Claim 3

The conjugate of claim 1 wherein the polypeptide is an antibody selected from the group consisting of naturally occurring monoclonal, polyclonal, chemically synethsized and recombinant DNA (rDNA)-derived.

Claim 4

The conjugate of claim 1 wherein the polypeptide is bonded to an intermediate "spacer arm" compound which is also bonded to the polymerizable compound.

Claim 5

The conjugate of claim 1 wherein the polymerizable compound is soluble in water or water/polar organic solvent mixtures.

Claim 6

The conjugate of claims 1 or 3 homopolymerized to form a polymer integrally containing the polypeptide.

Claim 7

The conjugate of claims 1 or 3 copolymerized with nonderivatized monomer to form a copolymer integrally containing the polypeptide.

Claim 8

The conjugate of claims 1 or 3 wherein the polymerizable compound is a compound containing olefinic or acetylenic unsaturation and at least one reactive site for bonding with the polypeptide or intermediate "spacer arm" compound.

Claim 9

The conjugate of claims 1 or 3 wherein the polymerizable compound is a polyunsaturated oligomer having at least one reactive site for bonding with the polypeptide or intermediate "spacer arm" compound.

Claim 10

The conjugate of claim 1, the polymerizable compound comprising a polymerizable, ethylenically unsaturated organic monomer selected from the group consisting of: where R l is hydrogen or a lower alkyl having from one to eight carbon atoms and R 2 is selected from the group consisting of -H -COC1 -COOH -C0 2 (CH 2 ) n OH( n = 1-8) -CH 2 NH2 -CH 2 Cl -C0 2 C 2 H 4 NHR (R = H or any organic group) -C0 2 CH 2 -CHOHCH 2 0H -CHO -C0 2 (CH 2 ) n NCO (n = 1-8) and R 3 and R 4 are selected from the group consisting of H and compounds which will provide an unsaturated group.

Claim 11

The conjugate of claim 9 wherein R 3 and R 4 are H.

Claim 14

The method of claim 11 wherein the substance to be removed is an antigen, the polypeptide is an antibody, and the fluid mixture is aqueous.

Claim 15

The method of claim 11, including providing a solid surface having at least one polymerizable compound bonded thereto capable of copolymerizing with the polymerizable organic compound/polypeptide conjugate, and polymerizing the conjugate in the presence of the polymerizable compound bonded to the solid surface.

Claim 16

An immunoassay method for determining the presence of an analyte in a fluid sample suspected of containing an analyte comprising: (a) contacting the fluid sample with a monomer/reactant conjugate in order to form a monomer/reactant conjugate--analyte complex and providing a reporter for labeling the monomer/reactant conjugate--analyte complex; (b) separating the labeled complex by initiating polymerization of the monomer/reactant conjugate--analyte complex; and (c) detecting the incorporation of reporter into the polymerized complex.

Claim 17

An immunoassay method for determining the presence of an analyte in a fluid sample suspected of containing an analyte comprising: (a) contacting the fluid sample with a monomer/reactant conjugate in order to form a monomer/reactant conjugate--analyte complex and providing a reporter for labeling the monomer/reactant conjugate--analyte complex; (b) separating the labeled complex by initiating polymerization of the monomer/reactant conjugate--analyte complex; and (c) detecting the incorporation of reporter into the polymerized complex.

Claim 18

The immunoassay method of claim 14 wherein one reporter is provided by a reporter/reactant conjugate capable of recognizing and binding to the analyte, the monomer/reactant conjugate or the monomer/reactant conjugate--analyte complex.

Claim 19

An immunoassay method for determining the presence of an analyte in a fluid sample suspected of containing the analyte comprising: (a) contacting the fluid sample with a monomer/analyte conjugate to form a fluid sample mixture; (b) contacting the mixture with a reporter/reactant conjugate capable of specifically binding analyte to form reporter-labeled analyte complex and reporter-labeled monomer/analyte complex; (c) separating the reporter-labeled monomer/analyte complex by initiating polymerization of the monomer/analyte-conjugate complex; and (d) detecting the incorporation of reporter into the polymerized complex.

Claim 20

An immunoassay method for determining the presence of an analyte in a fluid sample suspected of containing the analyte comprising: (a) contacting the fluid sample with a monomer/analyte conjugate to form a fluid sample mixture; (b) contacting the mixture with a reporter/reactant conjugate capable of specifically binding analyte to form reporter-labeled analyte complex and reporter-labeled monomer/analyte complex; (c) separating the reporter-labeled monomer/analyte complex by initiating polymerization of the monomer/analyte-conjugate complex; and (d) detecting the incorporation of reporter into the polymerized complex.

Claim 21

The immunoassay method of claim 14 or 16 wherein the reporter is selected from the group consisting of radioisotopes, enzymes, enzyme inhibitors, enzyme cofactors, fluorophores, luminescent materials, chromophores and particles.

Claim 22

The immunoassay method of claim 14 or 16, the monomer comprising a polymerizable, ethylenically unsaturated organic monomer selected from the group consisting of: where R 1 is a hydrogen or lower alkyl having from 1 to 8 carbon atoms, R 2 is selected from the group consisting of: -H -COC1 -COOH -C0 2 (CH 2 ) n OH (n = 1-8) -CH 2 NH2 -CH 2 Cl - C02C2HNHR (R = H or any organic group) -C0 2 CH 2 CHOHCH 2 0H -CHO - C02 (CH 2 ) n NCO (n = 1-8) and R 3 and R 4 are selected from the group consisting of H and compounds which will provide an unsaturated group.

Claim 23

The immunoassay method of claim 18 wherein R 3 and R 4 are H.

Claim 24

The immunoassay method of claim 14 or 16, the monomer comprising an acetylenically unsaturated monomer having a reactable group.

Claim 25

The immunoassay method of claim 14 or 16 wherein the monomer is 2-hydroxyethyl methacrylate.

Claim 26

The immunoassay method of claim 14 or 16 wherein the reporter is fluorescein isothiocyanate.

Claim 27

The immunoassay method of claim 14 or 16 wherein the reactants and analytes are selected from the group consisting of antibodies, antigens and haptens.

Claim 28

An immunoassay method for determining the presence of more than one analyte in a fluid sample suspected of containing one or more of said analytes, comprising: (a) contacting said fluid sample with respective monomer/reactant conjugates, one for each analyte thought to be present in the fluid sample to form respective monomer/reactant conjugate--analyte complexs and providing a reporter for labeling each monomer/reactant conjugate-analyte complex, each reporter providing a detectably different signal from every other reporter present in said fluid sample; (c) separating said respective labeled complexes by initiating polymerization of the respective monomer/reactant conjugate--analyte complexes; and (d) detecting the incorporation of each reporter into each said polymerized complex.

Claim 29

An immunoassay method for determining the presence of more than one analyte in a fluid sample suspected of containing one or more of said analytes, comprising: (a) contacting said fluid sample with respective monomer/reactant conjugates, one for each analyte thought to be present in the fluid sample to form respective monomer/reactant conjugate--analyte complexs and providing a reporter for labeling each monomer/reactant conjugate-analyte complex, each reporter providing a detectably different signal from every other reporter present in said fluid sample; (c) separating said respective labeled complexes by initiating polymerization of the respective monomer/reactant conjugate--analyte complexes; and (d) detecting the incorporation of each reporter into each said polymerized complex.

Claim 30

The immunoassay method of claim 24 wherein the reporter is selected from the group consisting of radioisotopes, enzymes, enzyme inhibitors, enzyme cofactors, fluorophores, luninescent materials, chromophores and particles.