Specific binding analytical and separation methods using carboxy containing polymers

Claim 3

The method of claim 1 wherein said ligand is a protein, carbohydrate, hapten, hormone or drug, and said receptor is an antibody for said protein, carbohydrate, hapten, hormone or drug.

Claim 4

The method of claim 1 wherein said ligand is biotin or a derivative thereof, and said receptor is avidin or a derivative thereof.

Claim 5

The method of claim 1 wherein said ligand is a single-stranded nucleic acid, and said receptor is a second single-stranded oligonucleotide which is substantially complementary to said nucleic acid.

Claim 6

A competitive binding assay for the determination of a specific binding ligand comprising: A. contacting a specimen suspected of containing a water-soluble specific binding ligand with a water-soluble receptor therefor, and with a reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR10## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) molecules of said ligand covalently attached to said particle through said reactive carboxy group or salt thereof, to form specific binding complex (i) between said receptor and said ligand, and specific binding complex (ii) between said receptor and said water-insoluble reagent, and B. after separating said complexes (i) and (ii), detecting the presence of either complex as an indication of the presence or amount of said ligand in said specimen.

Claim 7

A competitive binding assay for the determination of a specific binding ligand comprising: A. contacting a specimen suspected of containing a water-soluble specific binding ligand with a water-soluble receptor therefor, and with a reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR10## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) molecules of said ligand covalently attached to said particle through said reactive carboxy group or salt thereof, to form specific binding complex (i) between said receptor and said ligand, and specific binding complex (ii) between said receptor and said water-insoluble reagent, and B. after separating said complexes (i) and (ii), detecting the presence of either complex as an indication of the presence or amount of said ligand in said specimen.

Claim 10

The method of claim 6 wherein said immunological species is an antigenic material and said receptor is an antibody for said antigenic material.

Claim 11

The method of claim 6 wherein said immunological species is an antibody and said receptor is an antigenic material specific for said antibody.

Claim 12

The method of claim 6 wherein said immunological species is a first antibody and said receptor is a second antibody directed to said first antibody.

Claim 13

The method of claim 6 wherein said uncomplexed materials are separated from said immunological complex by filtration with a microporous membrane.

Claim 14

The method of claim 6 wherein said reagent is immobilized on a microporous filtration membrane.

Claim 15

The method of claim 6 wherein, either prior to, simultaneously with or subsequently to the formation of said water-insoluble immunological complex, said immunological species is reacted with a water-soluble specific binding component specifically reactive therefor.

Claim 16

The method of claim 12 wherein said specific binding component is labeled for detection.

Claim 17

The method of claim 13 wherein said label is an enzyme or radioisotope.

Claim 18

The method of claim 6 wherein said immunological species is human chorionic gonadotropin, Streptococcus A or a microorganism associated with periodontal disease.

Claim 19

An assay for the determination of a specific binding ligand comprising: detecting the presence or amount of a water-insoluble specific binding complex formed between a specific binding ligand of interest and a receptor for said ligand as an indication of the determination of said specific binding ligand of interest, the receptor provided as a component of a reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR12## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) the receptor for said ligand being covalently attached to said particle through said reactive carboxy group or salt thereof.

Claim 20

An assay for the determination of a specific binding ligand comprising: detecting the presence or amount of a water-insoluble specific binding complex formed between a specific binding ligand of interest and a receptor for said ligand as an indication of the determination of said specific binding ligand of interest, the receptor provided as a component of a reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR12## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) the receptor for said ligand being covalently attached to said particle through said reactive carboxy group or salt thereof.

Claim 21

An analytical separation method comprising: A. passing a specimen containing a mixture of biologically active substances over an affinity chromatography reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR13## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) a specific binding substance covalently attached to said particle through said reactive carboxy group, said specific binding substance being specific to one or more predetermined biologically active substances in said specimen mixture of biologically active substances to form a complex of said reagent with said predetermined substances, and B. collecting either the one or more complexed predetermined substances or one or more substances remaining in the eluent.

Claim 22

An analytical separation method comprising: A. passing a specimen containing a mixture of biologically active substances over an affinity chromatography reagent comprising: (I) a water-insoluble particle composed of a copolymer having recurring units derived from: (a) from about 60 to about 99.8 mole percent of one or more ethylenically unsaturated polymerizable oleophilic monomers which provide hydrophobicity to said copolymer, (b) from about 0.2 to about 40 mole percent of one or more ethylenically unsaturated polymerizable monomers having a reactive carboxy group, or salt thereof, and represented by the structure: ##STR13## wherein R is hydrogen, halo or alkyl of 1 to 3 carbon atoms, M is hydrogen, an alkali metal ion or an ammonium ion and L is an organic linking group having from 8 to 50 atoms selected from the group consisting of carbon, nitrogen, oxygen and sulfur atoms in the linking chain, said organic linking group further defined as having two or more divalent groups selected from the group consisting of alkylene, arylene, alkylenearylene and arylenealkylene which are connected to each other or terminated with an oxy, thio, imino, carbonyloxy, carbonylimino, ureylene or sulfonylimino group, and (c) from 0 to about 15 mole percent of one or more additional ethylenically unsaturated polymerizable monomers other than those identified in categories (a) and (b) above, and (II) a specific binding substance covalently attached to said particle through said reactive carboxy group, said specific binding substance being specific to one or more predetermined biologically active substances in said specimen mixture of biologically active substances to form a complex of said reagent with said predetermined substances, and B. collecting either the one or more complexed predetermined substances or one or more substances remaining in the eluent.