Use of Human IgM Derived Fragments to Study Structures Responsible for Protein A-Reactivity

Abstract
The protein A-binding site of human IgM was studied by affinity chromatography on SpA-Sepharose using fragments derived from a human monoclonal SpA-reactive IgM, Iz. Neither Fabμ nor (Fc)5μ fragments were retained on the column but IgM reactivity was unaffected by thermic treatment during proteolysis. Products intermediate between IgM and (Fc)5μ fragments produced during shorter proteolysis showed a reactivity related to their content in Fabμ regions. On the other hand mild reduction of IgM Iz to monomeric subunits results in a dramatic loss of SpA-affinity. However these subunits, like F(ab′)2μ but unlike Fab′μ fragments, showed a significant interaction with the column. Thus, the principal requirement for SpA reactivity with IgM Iz seems to be related to the presence of Fabμ regions in a polymeric state resembling native IgM.

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