Rapid purification of pertussis toxin (PT) and filamentous hemagglutinin (FHA) by cation-exchange chromatography.
Pertussis toxin (PT) and filamentous hemagglutinin (FHA) were purified from culture supernatant of Bordetella pertussis Saadet and Tohama strains, using CM-Sepharose CL-6B cation-exchange chromatography. By the rapid purification method described here, both proteins were separately eluted from the same column in pure forms. The PT and FHA in the extract of culture supernatant were bounded to CM-Sepharose CL-6B cation-exchange column in 50 mM phosphate buffer containing 2 M urea (Buffer A), pH 6.0. Then the PT was eluted from the column with Buffer A (pH 7.4) and after elution of the PT, the FHA was eluted with 0.5 M NaCl in 50 mM phosphate buffer. Pertussis toxin and filamentous haemagglutinin purified by this procedure were electrophoretically and immunologically identical to the reference preparations.
Ozcengiz
Erkan
E
Bacterial Vaccines Research Laboratory, Refik Saydam Hygiene Center, 06100 Ankara, Turkey.
Kilinç
Kamer
K
Büyüktanir
Ozlem
O
Günalp
Ayfer
A
D000818:Animals; D001886:Bordetella pertussis / Q000254:growth & development / Q000378:metabolism; D002645:Chickens; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000276:immunology; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000302:isolation & purification*; D006632:Histamine / Q000494:pharmacology; D066298:In Vitro Techniques; D007964:Leukocytosis; D051379:Mice; D037342:Pertussis Toxin / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives*
D006388:Hemagglutinins; C047264:sepharose CL 6B; D006632:Histamine; D012685:Sepharose; D037342:Pertussis Toxin