39] Solid-phase radioimmunoassay for immunoglobulins and influenza antibodies

Abstract

Publisher Summary This chapter discusses the solid-phase radioimmunoassay for immunoglobulins and influenza antibodies. The conventional serological techniques have been innovated by the development of various solid-phase immunoassays. These procedures have made serological testing more precise, rapid, and easier, and they have permitted greater experimental versatility in characterization and purification of viral antigens and antibodies. Heavy chains were prepared by dialyzing the chromatographic IgG eluate against 0.3 M Tris(hydroxy-methyl)aminomethane(Tris) hydrochloride buffer at pH 8.5. The IgG was then reduced with 0.3 M 2-mercaptoethanol and the heavy- and light-chain mixture was alkylated with equimolar iodoacetate. The indirect viral antibody assay was used, incorporating various influenza viral strains and antibodies. The direct quantitative relationship for variations in viral antibody concentration and binding of the radiolabeled indicator was observed. The direct and indirect methods of RIA were compared for the minimal amounts of viral antibody or Ig detected. End-point dilutions were the lowest concentration of viral antibody or Ig that resulted in a plateau of the binding of indicator (anti-Ig) in the indirect assays or the final labeled reactant in direct assays.