Removal of LPS from a Brucella cytoplasmic fraction by affinity chromatography with an anti-LPS monoclonal antibody as immunosorbent.
Affinity chromatography on polymyxin B-Sepharose 4B is one of the most commonly used methods for the removal of contaminating lipopolysaccharides (LPS). However, the LPS of Brucella spp. do not bind to polymyxin B. An affinity chromatography method with an anti-O antigen of Brucella LPS monoclonal antibody as immunosorbent was developed. The method produced a 1000-fold reduction in the LPS content of the cytoplasmic fraction of B. abortus. The eluted proteins retained their antigenicity. The method, which uses mild physiological conditions, is simple, effective and reproducible.
Goldbaum
F A
FA
Instituto de Estudios de la Inmunidad Humoral (IDEHU) (UBA-CONICET), Facultad de Farmacia y Bioquimica, Buenos Aires, Argentina.
Rubbi
C P
CP
Fossati
C A
CA
D000372:Agglutination Tests; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D002003:Brucella abortus / Q000737:chemistry* / Q000648:ultrastructure; D002846:Chromatography, Affinity / Q000379:methods*; D003593:Cytoplasm / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D015151:Immunoblotting; D005779:Immunodiffusion; D007164:Immunosorbents / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology / Q000302:isolation & purification*; D019081:O Antigens; D011135:Polysaccharides, Bacterial / Q000276:immunology; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007164:Immunosorbents; D008070:Lipopolysaccharides; D019081:O Antigens; D011135:Polysaccharides, Bacterial