The Antigens of Mycobacterium bovis, Strain BCG, Studied by Crossed Immunoelectrophoresis: a Reference System

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Abstract
A reference system for the soluble antigens of M. bovis, strain BCG, is described. The system is based on crossed immunoelectrophoresis with intermediate gel. A commercially available Ig preparation made from rabbit anti-BCG hyperimmune serum was used as reference antibodies; a concentrated BCG culture filtrate was used as reference antigen. The pattern obtained was highly reproducible; most of the components were stable when the 50-fold-concentrated culture filtrate was stored at -20.degree. C. About 30 different antigenic components were selected as reference antigens and numbered. The majority of the reference antigens were present in extracts prepared from BCG by ultrasonication or bacterial press extraction. Use of the system for studies of antigenic relationship between mycobacteria, identification and quantification of antigens and characterization of antimycobacterial antibodies are illustrated by examples. The antigens of 2 preparations of tuberculin purified protein derivative (PPD) were identified. The antigen designated BCG60 was a major constituent of tuberculin PPD. Evidence is presented that this antigen is cell wall associated.

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