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. 2013 Oct 1:102:2.5.1-2.5.29.
doi: 10.1002/0471142735.im0205s102.

Production of monoclonal antibodies

Wayne M Yokoyama  1 , Michelle Christensen  2 , Gary Dos Santos  2 , Diane Miller  2 , Jason Ho  3 , Tao Wu  3 , Michael Dziegelewski  3 , Francisca A Neethling  3
Affiliations

Production of monoclonal antibodies

Wayne M Yokoyama et al. Curr Protoc Immunol. .

Abstract

This unit describes the production of monoclonal antibodies beginning with immunization, cell fusion, and selection. Support protocols are provided for screening primary hybridoma supernatants for antibodies of desired specificity, establishment of stable hybridoma lines, cloning of these B cell lines by limiting dilution to obtain monoclonal lines, and preparation of cloning/expansion medium. An alternate protocol describes cell fusion and one-step selection and cloning of hybridomas utilizing a semi-solid methylcellulose-based medium (ClonaCell-HY from StemCell Technologies).

Keywords: B cell hybridomas; limiting dilution; monoclonal antibodies; semi-solid methylcellulose-based medium.

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References

Literature Cited

References
    1. Bazin, R. and Lemieux, R. 1989. Increased proportion of B cell hybridomas secreting monoclonal antibodies of desired specificity in cultures containing macrophage-derived hybridoma growth factor (IL-6). J. Immunol. Methods 116:245-249.
    1. Coffino, P., Baumal, R., Laskov, R., and Scharff, M.D. 1972. Cloning of mouse myeloma cells and detection of rare variants. J. Cell. Physiol. 79:429-440.
    1. Davis, J.M., Pennington, J.E., Kubler, A-M., and Conscience, J.F. 1982. A simple, single-step technique for selecting and cloning hybridomas for the production of monoclonal antibodies. J. Immunol. Methods 50:161-171.
    1. Goding, J.W. 1986. Monoclonal Antibodies: Principles and Practice. Academic Press, San Diego.
    1. Harlow, E. and Lane, D. 1988. Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
Key References
    1. Goding, 1986. See above.
    1. Köhler and Milstein, 1975. See above.
    1. Oi, V.T. and Herzenberg, L.A. 1980. Immuno-globulin-producing hybrid cell lines. In Selected Methods in Cellular Immunology (B.B. Mishell and S.M. Shiigi, eds.) pp. 351-372. W.H. Freeman, New York.
Internet Resources
    1. http://www.stemcell.com/en/Products/All-Products/ClonaCellHY-Hybridoma-K...

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