Antigenic mosaic of Methanosarcinaceae: partial characterization of Methanosarcina barkeri 227 surface antigens by monoclonal antibodies

Abstract

Hybridomas were constructed with spleen cells from mice immunized against Methanosarcina barkeri 227. The reaction with the resulting monoclonal antibodies identified two antigenic determinants. Determinant 8A is present in M. barkeri 227, where it is accessible to antibody on whole bacterial cells. 8A is undetectable in (or absent from) M. barkeri R1M3, an immunologically closely related strain. Determinant 8C is present in both strains, but with M. barkeri 227 it is found only in extracts and cannot be demonstrated in whole cells. It therefore appears to be hidden. A soluble form of antigen 8A (antigen 227) was obtained treating whole M. barkeri 227 cells with absolute methanol. This antigen was further purified by affinity chromatography with antibody 8A. Chemical and immunochemical analyses of these preparations showed that antigen 227 is a high-molecular-weight (4 X 10(5)) structure composed mainly of one carbohydrate, glucose, and small amounts of amino acids. Its solubility properties suggest that this molecule is associated with a lipid moiety.