Influence of molecular size of IgA on its immunoassay by various techniques. II. Solid-phase radioimmunoassays

Abstract

Homogeneous, polymeric (P), monoclonal (MC) and polyclonal (PC) samples of purified IgA, as well as of secretory IgA (sIgA), were compared to their corresponding monomeric (M) forms with regard to their respective behaviour in both a solid-phase double antibody (AB) immunoradiometric assay (IRMA) and a solid-phase competition radioimmunoassay (CRIA). On an identical weight basis, both assays underestimated the P forms. Correction factors (CF), i.e., optical density (OD) versus radiometric concentration ratios, were calculated for both methods for all P forms, using M as standards. IRMA CF were respectively 1.50 for dimer, 1.98 for secretory IgA and 2.40 for tetramers, very similar to those obtained in radial immunodiffusion (RID). With optimally coated beads, these CF were stable along a wide range of concentrations. In contrast, CRIA-CF were 3-4 times larger and displayed much variation along their standard range of concentrations, preventing the use of a constant CF along the standard curve. Our present IRMA, with its CF, should be of value for a more accurate quantitation of small amounts of IgA of various known sizes.