Abstract |
The heat-treated apoceruloplasmin (Apocp) is a useful protein as an affinity ligand for the purification of pertussis toxin (PT). The amounts of Apocp in the purified antigens or the pertussis component vaccine were determined. Anti-Apocp antibodies were not detected by the passive cutaneous anaphylaxis (PCA) test in rats. No anti-Apocp antibody was detected after hyperimmunization of rabbits with the vaccine. Apocp was not detected in PT and filamentous hemagglutinin (FHA) by ELISA using rabbit anti-Apocp IgG. In the experiments using 125I-labelled Apocp, 125I-Apocp was not detected in either PT or FHA which were purified by 125I-labeled Apocp- Sepharose, DEAE Sepharose, and cellulose sulfate chromatography. The contents of human DNA were also determined to be less than 10 pg per 1 mg of Apocp, by the dot-blot hybridization method using the 32P-labeled DNA probe of Alu sequence. In the tests for the presence of inapparent viruses, HBs antigen and HTLV-III antibody, no contamination was found in either the Apocp or in the vaccine. Large amounts of various viruses, which were intentionally added to the Apocp (spiking test), were completely inactivated by heating at 65 degrees C for 18 hr. Both the Apocp and the vaccine passed the general pharmacology and acute toxicity tests. From these results, the heat-treated Apocp was considered to be a suitable affinity ligand for the purification of the antigens for the pertussis component vaccine.
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Authors | A Ginnaga, K Morokuma, S Furukawa, T Shigaki, N Katsuki, K Aihara, T Oka, M Sakoh, C Tamura, A Imaizumi |
Journal | Developments in biological standardization
(Dev Biol Stand)
Vol. 73
Pg. 233-41
( 1991)
ISSN: 0301-5149 [Print] Switzerland |
PMID | 1778315
(Publication Type: Journal Article)
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Chemical References |
- Adhesins, Bacterial
- Apoproteins
- Hemagglutinins
- Pertussis Vaccine
- Virulence Factors, Bordetella
- apoceruloplasmin
- filamentous hemagglutinin adhesin, Bordetella pertussis
- DNA
- Ceruloplasmin
- Pertussis Toxin
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Topics |
- Adhesins, Bacterial
- Animals
- Apoproteins
(analysis, toxicity)
- Ceruloplasmin
(analysis, toxicity)
- Chromatography, Affinity
- DNA
(analysis)
- Evaluation Studies as Topic
- Hemagglutinins
(analysis)
- Hot Temperature
- Humans
- Mice
- Mice, Inbred ICR
- Pertussis Toxin
- Pertussis Vaccine
(analysis, isolation & purification)
- Safety
- Virulence Factors, Bordetella
(analysis)
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