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AFFINITY CHROMATOGRAPHY FOR PURIFICATION OF ANTIBODIES TO NEISSERIA GONORRHOEAE AND NEISSERIA MENINGITIDIS LIPOPOLYSACCHARIDES
Author(s) -
RDAHL EYVIND,
MÆLAND JOHAN A.
Publication year - 1984
Publication title -
acta pathologica microbiologica scandinavica series c: immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0108-0202
DOI - 10.1111/j.1699-0463.1984.tb00083.x
Subject(s) - affinity chromatography , neisseria gonorrhoeae , antibody , pronase , chemistry , sepharose , chromatography , neisseria , antigen , microbiology and biotechnology , bacteria , trypsin , biology , biochemistry , enzyme , immunology , genetics
Lipopolysaccharides (LPSs) were prepared by phenol‐water extraction of the gonococcal strain 8551 and the group B meningococcal strain 44/76, digested with pronase, and purified by ultracentrifugation and Sepharose CL‐6B fractionation in the presence of 1.5 per cent sodium deoxycholate. On SDS‐PAGE with 10 per cent acrylamide the purified 5I‐labelled LPSs migrated as single, low‐molecular‐weight components. The LPSs were coupled to CNBr‐activated Sepharose 4B for affinity purification of antibodies to the common antigenic factor 1 and the sero‐type factor 5 of LPS 8551, and antibodies to LPS 44/76. The antibodies eluted showed ELISA activity against wells coated with LPS or whole cells of the bacteria, the antibody activity being inhibited by LPS. SDS‐PAGE of whole cells of the strain 8551 and immunoblotting with the anti‐factor 1 or ‐factor 5 antibodies resulted in single, broad bands corresponding to the low‐molecular‐weight LPS subunits.