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 s...

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Bibliographic Details
Published in:Vaccine 2004-03, Vol.22 (11), p.1570-1575
Main Authors: Ozcengiz, Erkan, Kilinç, Kamer, Büyüktanir, Ozlem, Günalp, Ayfer
Format: Article
Language:English
Subjects:
Animals
Applied microbiology
Biological and medical sciences
Bordetella pertussis
Bordetella pertussis - growth & development
Bordetella pertussis - metabolism
Chickens
Chromatography
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Enzymes
Erythrocytes - immunology
Filamentous hemagglutinin
Fundamental and applied biological sciences. Psychology
Hemagglutination Inhibition Tests
Hemagglutinins - isolation & purification
Histamine - pharmacology
In Vitro Techniques
Leukocytosis
Methods
Mice
Microbiology
Molecular weight
Pertussis toxin
Pertussis Toxin - isolation & purification
Proteins
Purification
Sepharose - analogs & derivatives
Sodium chloride
Studies
Toxins
Urea
Vaccine
Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)
Whooping cough
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Summary: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.
ISSN:0264-410X
1873-2518
DOI:10.1016/j.vaccine.2003.09.040