High yield monoclonal antibody production in ascites

Brodeur, Bernard R.; Tsang, Peggy S.

doi:10.1016/0022-1759(86)90459-x

Cited by 33 publications

(10 citation statements)

References 3 publications

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“…B. pertussis antibody-positive cells were subcloned by limiting dilution. Selected subclones were grown for antibody production in serum-free cell culture medium, or as ascites as previously described (Brodeur & Tsang, 1986). mAbs were purified as previously described (Hamel & Brodeur, 1990).…”

Section: Methodsmentioning

confidence: 99%

Characterization and comparative bactericidal activity of monoclonal antibodies to Bordetella pertussis lipo-oligosaccharide A

Archambault

Rondeau

Martin

et al. 1991

Journal of General Microbiology

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Spleen cells from mice immunized with a Bordetefla pertussis N-lauroyl sarcosine membrane extract (SME) were used to generate hybridoma cells lines producing monoclonal antibodies (mAbs). Seven mAbs were shown to be specific to B. pertussis lipo-oligosaccharide (LOS) by immunoblotting of SME or purified LOS following SDS-PAGE. All mAbs reacted with the B. pertussis Tohama I strain of the LOS AB phenotype, and did not react with the atypical variant strain 134 of the LOS B phenotype. The immune reactivity of the mAbs was retained after treatment of SME with proteinase K and was lost after sodium periodate treatment. No cross-reactivity was observed with the mAbs when tested against B. parapertussis and other Gram-negative bacteria. However, all mAbs reacted with B. bronchiseptica. Binding assays with live B. pertussis cells demonstrated that mAbs strongly reacted with cell surface exposed antigenic determinants. High bacterial cell lytic capability was observed for five of these mAbs. Concentrations between 0.22 and 2.2 pg mAb ml-l (0.1 and 1 pg per 450 p1 assay) purified by protein A were required to kill at least 50% of the bacteria. Competition immunoassays with biotinylated antibodies showed that the bacteriolytic and non-bacteriolytic mAbs were directed to different epitopes of the B. pertussis LOS A.

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Section: Methodsmentioning

confidence: 99%

Characterization and comparative bactericidal activity of monoclonal antibodies to Bordetella pertussis lipo-oligosaccharide A

Archambault

Rondeau

Martin

et al. 1991

Journal of General Microbiology

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“…http://iacuc.cwru.edu Pistillo et al 1992Brodeur and Tsang 1986Stewart et al 1989 Hybridoma cells Type of cell Some cells show a large variety in growth pattern in animals and tend to grow poorly in some animals. Some cells have an aggressive growth pattern and are often poor MAb producers.…”

Section: Mab Production: Guidelinesmentioning

confidence: 99%

Critical Steps in the Production of Polyclonal and Monoclonal Antibodies: Evaluation and Recommendations

Leenaars¹,

Hendriksen²

2005

ILAR Journal

173

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Antibodies are valuable tools in the laboratory and clinic. Antibodies include those secreted by a single clone of B lymphocytes, termed monoclonal antibodies, and those produced by a mixture of various B lymphocyte clones, termed polyclonal antibodies. Both products have become essential instruments in fundamental immunological research, immunohistochemistry, diagnostic testing, and vaccine quality control. Antibody production requires a substantial number of animals, and the animals are subjected to a number of invasive procedures such as antigen injection and blood collection. However, by carefully designing an immunization protocol and by optimizing the immunization response, it is possible to minimize animals' pain and distress while obtaining optimal immune responses. In this article, the critical steps in the production of polyclonal and monoclonal antibodies are described, specifically including selection of the animal species and its age, injection protocol, and ascites tapping. Recommendations are provided for optimizing the immunization response.

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“…At the conclusion of these limiting dilutions, the selected clones were expanded for freezing and ascites fluid production. The ascites fluid was produced in F1 mouse hybrids (Swiss Webster X BALB/c) according to previously described procedures, (16,17).…”

Section: Fusion Procedures and Monoclonal Antibody Productionmentioning

confidence: 99%

Neutralizing Monoclonal Antibody Against Enterovirus-70 Reacts with Viral Proteins 1C and ID

Wiley¹,

Brodeur²,

Dimock³

et al. 1990

Viral Immunology

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A library of murine monoclonal antibodies against the prototype Enterovirus-70 (EV-70) strain, J670/71, was made for the purpose of studying the immunologically reactive determinants of the virus. Each of the monoclonal antibodies reacted with several other strains of Enterovirus-70 when tested by immunofluorescence. However, none of these monoclonal antibodies reacted with any other picornavirus tested. It was found that all of the monoclonal antibodies precipitated EV-70 viral proteins 1C and 1D in radio-immunoprecipitation assays. However, only one of these monoclonal antibodies, an IgG3 kappa, was capable of neutralizing the virus.

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High yield monoclonal antibody production in ascites

Cited by 33 publications

References 3 publications

Characterization and comparative bactericidal activity of monoclonal antibodies to Bordetella pertussis lipo-oligosaccharide A

Characterization and comparative bactericidal activity of monoclonal antibodies to Bordetella pertussis lipo-oligosaccharide A

Critical Steps in the Production of Polyclonal and Monoclonal Antibodies: Evaluation and Recommendations

Neutralizing Monoclonal Antibody Against Enterovirus-70 Reacts with Viral Proteins 1C and ID

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