Enzyme linked immunoassay with stabilized polymer saccharide enzyme conjugates

In an immunoassay method of detection wherein the immunoassay method comprises contacting an antibody polymer conjugate with an antigen so as to bind the antigen, and measuring the amount of antigen bound to the antibody polymer conjugate, the improvement wherein the antibody polymer conjugate comprises antibody covalently bonded to a polymer by linker groups, wherein the polymer is selected from the group consisting of a polyvinyl polymer, a polyester polymer, a polyamide polymer and mixtures thereof, the polymer having a main chain molecular weight of at least 5,000, and wherein three or more linker groups covalently bond both the antibody and polymer and said linker groups have at least three hydroxyl groups and not more than 60 carbon atoms, each of said linker groups being selected from the group consisting of ##STR25## wherein E=--OH or --SH;

• n=0 or 1;
• W=0 or 1;
• and
• Y=bond or --CH.sub.2 ;
• wherein A is bonded to the polymer and is selected from the group consisting of ##STR26## B is bonded to the antibody and is selected from the group consisting of ##STR27## wherein R.sub.1 and R.sub.2 are each selected from the group consisting of H, OH, acetoxy, alkoxy, phenylene, straight chain alkyl or branched chain alkyl having 1 to 12 carbons, ##STR28## wherein x=2 to 7, ##STR29## 2-hydroxy ethyl, 3-chloro-2-hydroxypropyl, 2-hydroxy-1-methylethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2, 3-dihydroxypropyl, 2 hydroxybutyl, 4 -hydroxybutyl, diethylene-glycol, 5-hydroxypentryl, 6-hydroxyhexyl, triethyleneglycol, 7-hydroxyheptyl, 3,4-dihydroxybutyl, 5-hydroxypentyl, 6-hydroxyhexyl, 2,3-dimethyl-3-hydroxybutyl, 5,6-dihydroxyhexyl, 7-hydroxyheptyl, a residue of a mono-, di-, and trisaccharide;
• wherein R.sub.3 selected from the group consisting of H, straight chain alkyl or branched chain alkyl having 1 to 12 carbons, ##STR30## wherein x=2 to 7, ##STR31## 2-hydroxy ethyl, 3-chloro-2-hydroxypropyl, 2-hydroxy-1-methylethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2, 3-dihydroxypropyl, 2 hydroxybutyl, 4-hydroxybutyl, diethylene-glycol, 5-hydroxypentryl, 6-hydroxyhexyl, triethyleneglycol, 7-hydroxyheptyl, 3,4-dihydroxybutyl, 5-hydroxypentyl, 6-hydroxyhexyl, 2,3-dimethyl-3-hydroxybutyl, 5,6-dihydroxyhexyl, 7-hydroxyheptyl, a residue of a mono-, di-, and trisaccharide;
• and
• R'.sub.1, R'.sub.2, R'.sub.3, R'.sub.4, R'.sub.5, R'.sub.6, R'.sub.7, R'.sub.8, and R'.sub.9 each selected from the group consisting of H, OH, acetoxy, alkoxy, halogen and phosphate, straight chain alkyl or branched chain alkyl having 1 to 12 carbons, ##STR32## wherein x=2 to 7, ##STR33## 2-hydroxy ethyl, 3-chloro-2-hydroxypropyl, 2-hydroxy-1-methylethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 2 hydroxybutyl, 4-hydroxybutyl, diethylene-glycol, 5-hydroxypentryl, 6-hydroxyhexyl, triethyleneglycol, 7-hydroxyheptyl, 3,4-dihydroxybutyl, 5-hydroxypentyl, 6-hydroxyhexyl, 2,3-dimethyl-3-hydroxybutyl, 5,6-dihydroxyhexyl, 7-hydroxyheptyl, a residue of a mono-, di-, and trisaccharide.