Method for solid phase immunological assay of antigen

Abstract

Abstract An improved solid-phase radioimmunological assay method for assay of an antigen in a biological fluid, e.g., blood plasma, by use of a solid phase polymeric substrate on which is adsorbed in solid phase an antibody composite layer, comprising a solid phase precoat of an anti-antibody (Ab 2 ) (i.e., an antibody to an animal blood serum and sometimes referred to as a second antibody) bound to said substrate and a solid phase antibody Ab specific to said antigen (sometimes referred to as a primary antibody) immunologically bound to said anti-antibody precoat. The biological fluid to be tested, containing the antigen and radioactive labelled antigen, is contacted with the antibody surface of the composite layer to immunologically bind antigen and radioactive labelled antigen to the solid phase Ab surface, whereupon the quantity of antigen in the biological fluid is determined by measuring the radioactivity of the composite layer and substrate or of the remaining biological fluid containing unbound antigen and unbound radioactive labelled antigen. An improved method for making as an article of manufacture solid phase antibody (Ab) for radioimmunological assay of antigen by first adsorbing Ab 2 from its serum on a solid substrate to produce a solid Ab 2 precoat and then reacting the Ab 2 precoat with Ab in its antiserum to form a solid phase Ab coating on the Ab 2 precoat whereby an effective, efficient, reproduceable and stable solid phase antibody Ab for assay of antigen is obtained with a substantial decrease in the amount of Ab required. The terms coating, precoating, coat and coating do not necessarily mean continuous or complete coat or coating of the underlying surface but, instead, include continuous and non-continuous coatings.

Inventors

• 👤 Gerald J. Litt

Bibliographic Details