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Quantitation and purification of the Pasteurella multocida toxin by using monoclonal antibodies. 1988

N T Foged
National Veterinary Laboratory, Copenhagen, Denmark.

Pasteurella multocida toxin (PMT), derived from a toxigenic strain of P. multocida originally isolated from a pig with clinical atrophic rhinitis, was used to immunize BALB/c mice. Ninety-two hybridomas secreting monoclonal antibodies (MAbs) against PMT were produced by fusion of spleen cells from these mice with P3-X63-Ag8.653 myeloma cells. The specificity for PMT of the MAbs was ascertained by enzyme-linked immunosorbent assay and immunoblotting analysis. The interrelationship of a panel of 10 MAbs and their respective epitopes was characterized by a competitive enzyme-linked immunosorbent assay based on the biotin-avidin system and by an in vitro neutralization assay based on the cytopathic effect of PMT on embryonic bovine lung cells. In vivo neutralization of the lethal effect of PMT in mice was obtained by passive immunization with an anti-PMT MAb 2 days before challenge with PMT. PMT was quantified by a sandwich enzyme-linked immunosorbent assay with a lower detection limit of approximately 50 pg of PMT. Application of supernatant or bacterial extract from cultivation of toxigenic P. multocida to an affinity column containing immobilized MAb resulted in purification of PMT with a yield of 78 to 93% of the PMT applied.

UI MeSH Term Description Entries
D007163 Immunosorbent Techniques Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody. Immunoadsorbent Techniques,Immunoadsorbent Technics,Immunosorbent Technics,Immunoadsorbent Technic,Immunoadsorbent Technique,Immunosorbent Technic,Immunosorbent Technique,Technic, Immunoadsorbent,Technic, Immunosorbent,Technics, Immunoadsorbent,Technics, Immunosorbent,Technique, Immunoadsorbent,Technique, Immunosorbent,Techniques, Immunoadsorbent,Techniques, Immunosorbent
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D009500 Neutralization Tests The measurement of infection-blocking titer of ANTISERA by testing a series of dilutions for a given virus-antiserum interaction end-point, which is generally the dilution at which tissue cultures inoculated with the serum-virus mixtures demonstrate cytopathology (CPE) or the dilution at which 50% of test animals injected with serum-virus mixtures show infectivity (ID50) or die (LD50). Neutralization Test,Test, Neutralization,Tests, Neutralization
D010325 Pasteurella The oldest recognized genus of the family PASTEURELLACEAE. It consists of several species. Its organisms occur most frequently as coccobacillus or rod-shaped and are gram-negative, nonmotile, facultative anaerobes. Species of this genus are found in both animals and humans.
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000907 Antibodies, Bacterial Immunoglobulins produced in a response to BACTERIAL ANTIGENS. Bacterial Antibodies
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000942 Antigens, Bacterial Substances elaborated by bacteria that have antigenic activity. Bacterial Antigen,Bacterial Antigens,Antigen, Bacterial
D001427 Bacterial Toxins Toxic substances formed in or elaborated by bacteria; they are usually proteins with high molecular weight and antigenicity; some are used as antibiotics and some to skin test for the presence of or susceptibility to certain diseases. Bacterial Toxin,Toxins, Bacterial,Toxin, Bacterial

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