Isolation of the lymphocytosis promoting factor-haemagglutinin of Bordetella pertussis by affinity chromatography.
The lymphocytosis promoting factor-haemagglutinin of Bordetella pertussis was isolated from solutions obtained after cell disintegration by a novel affinity chromatographic method using an adsorbent composed of human haptoglobin covalently attached to a Sepharose 4B matrix. The haemagglutinin was bound to the adsorbent at pH 6.5 and eluted by a stepwise change to a pH 10 buffer. A 300--600-fold purification of the haemagglutinin was achieved by this one-step process. The chemical and biological properties of the haemagglutinin isolated by affinity chromatography were found to be similar to those of the protein isolated by other workers from culture supernatants. The affinity chromatographic method was found to be specific for the purification of the lymphocytosis promoting factor-haemagglutinin and no purification of the fimbrial-haemagglutinin of Bordetella pertussis was achieved by the method.
D000373:Agglutinins / Q000302:isolation & purification*; D000818:Animals; D001427:Bacterial Toxins; D001886:Bordetella pertussis / Q000032:analysis* / Q000254:growth & development; D002846:Chromatography, Affinity; D006388:Hemagglutinins / Q000302:isolation & purification*; D006632:Histamine / Q000494:pharmacology; D007376:Interleukin-2 / Q000302:isolation & purification*; D007958:Leukocyte Count; D008222:Lymphokines / Q000302:isolation & purification*; D051379:Mice; D037342:Pertussis Toxin; D010566:Virulence Factors, Bordetella
D000373:Agglutinins; D001427:Bacterial Toxins; D006388:Hemagglutinins; D007376:Interleukin-2; D008222:Lymphokines; D010566:Virulence Factors, Bordetella; D006632:Histamine; D037342:Pertussis Toxin