Influence of molecular size of IgA on its immunoassay by various techniques—I. Direct and reversed single radial immunodiffusion

Abstract

Immunochemically pure samples of monoclonal and polyclonal IgA were prepared from human sera and milk. Samples of various homogeneous molecular sizes were further obtained by preparative ultracentrifugations. The different behaviour of each preparation (monomer, dimer, trimer, tetramer and secretory IgA) were studied in direct (DRID) and reversed (RRID) single radial immuno-diffusion using three different anti-α-chain antisera and IgA samples of various monoclonal and polyclonal origins. In DRID, all polymer concentrations were underestimated when using monomers as standards, on an equal weight (OD) basis. Correction factors (CFs) were calculated from monomer to polymer DRID slope ratios. The means ± SDs of these CFs were 1.55 ± 0.18 for serum dimers, 1.85 ± 0.10 for trimers, 2.63 ± 0.26 for tetramers and 2.24 ± 0.15 for secretory IgA (84% 11S, 16% 15S). These results were confirmed by RRID.