A novel process for preparing an acellular pertussis vaccine composed of non-pyrogenic toxoids of pertussis toxin and filamentous hemagglutinin.
A novel process for preparing non-pyrogenic toxoids of pertussis toxin (PT) and filamentous hemagglutinin (FHA) is described. The process consists of chromatographies on perlite then on hydroxylapatite. Purification yields for PT and FHA are 62 and 68%, respectively. The purification process takes advantage of the novel use of perlite (a filter aid) for the simultaneous purification of PT and FHA. The hydroxylapatite, in addition to removing the remaining contaminants, also concentrates the antigens. The resulting PT and FHA are approximately 95% pure, and are non-pyrogenic as judged by the rabbit pyrogen test. The purification process is simple, inexpensive, and does not use blood components or toxic substances. The mild conditions in which the PT and FHA are purified ensure the recovery of native protein. The purified PT and FHA are detoxified in the presence of glycerol using glutaraldehyde and formaldehyde, respectively, to produce antigenic components of an acellular pertussis vaccine. The final PT and FHA toxoids are immunogenic in guinea-pigs and have been shown to be protective in the mouse intracerebral challenge test.
Tan
L U
LU
Connaught Laboratories Limited, Willodale, Ontario, Canada.
Fahim
R E
RE
Jackson
G
G
Phillips
K
K
Wah
P
P
Alkema
D
D
Zobrist
G
G
Herbert
A
A
Boux
L
L
Chong
P
P
D001886:Bordetella pertussis / Q000032:analysis / Q000276:immunology; D006388:Hemagglutinins / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D037342:Pertussis Toxin; D010567:Pertussis Vaccine / Q000276:immunology*; D011749:Pyrogens / Q000032:analysis; D010566:Virulence Factors, Bordetella / Q000276:immunology / Q000302:isolation & purification*
D006388:Hemagglutinins; D010567:Pertussis Vaccine; D011749:Pyrogens; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin