Abstract
An indirect solid-phase radioimmunoassay involving purified influenza haemagglutinin bound to polyvinyl microtitre plates has been used to quantitate the humoral response to influenza infection in ferrets. Sialic acid residues, present on normal ferret immunoglobulin, caused non-specific binding of immunoglobulin to the haemagglutinin, resulting in a non-linear relationship between serum dilution and primary antibody bound to antigen. Direct proportionality was achieved by removal of sialic acid residues from serum immunoglobulin or by inclusion of free sialic acid in the incubation buffer. Absolute quantitation of IgG and IgM antibodies was achieved by determining the relationship between primary antibody bound to antigen and secondary anti-immunoglobulin antibodies using 131I-IgG or -IgM and the homologous 25I-labelled monospecific antiserum. Examples of antibody quantitations in sequential post-infection sera are presented.
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