Reversed immunosorbents: a simple method for specific antibody immobilization

A method is presented for permanently converting an antigen immunosorbent into a purified antibody immunosorbent which retains specific reactivity for antigen. Following immunological reaction of the antibodies in an antiserum with the antigen adsorbent, the specific immunoglobulin is chemically bound to the antigen by means of the divalent crosslinking agent. Various such agents, including glutaraldehyde, suberimidate, a diazide, and isocyanates, were employed. The conditions of preparation using the first two were optimized, and the characteristics of the resulting immobilized antibodies were investigated. Applications in radioimmunoassay, preparative chromatography, and affinity-constant studies discussed in view of the unique attributes of these reagents.