Abstract
Whereas the expression of foreign genes in mammalian cells usually proves successful, the purification of gene products is often a difficult and time-consuming process. The availability of monoclonal antibodies to the foreign protein can greatly assist in small scale purification, but where antibodies are not available, alternatives have to be sought One useful approach involves the fusion of the foreign gene adjacent to a gene segment encoding an antibody heavy chain variable region (1). By transfection of this construct into a cell line producing a compatible light chain or by cotransfection of the fusion product with a light chain gene, an antibody-like molecule can be produced and purified using the corresponding antigen.
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References
Neuberger, M. S., Williams, G. T., and Fox, R. O. (1984) Recombinant antibodies possessing novel effector functions. Nature 312, 604–608.
Williams, G. T. and Neuberger, M. S. (1986) Production of antibody-tagged enzymes by myeloma cells: Application to DNA polymerase I Klenow fragment. Gene 43, 319–324.
Mulligan, R. C. and Berg, P. (1981) Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc. Natl. Acad. Sci. USA 78, 2072–2076.
Lundblad, A., Steller, R., Kabat, E. A., Hirst, J. W., Weigert, M. G., and Cohn, M. (1972) Immunochemical studies on mouse myeloma proteins with specificity for dextran or for levan. Immunochemistry 9, 535–544.
Oi, V. T., Morrison, S. L., Herzenberg, L. A., and Berg, P. (1983) Immunoglobulin gene expression in transformed lymphoid cells. Proc. Nat. Acad. Sci. USA 80, 825–829.
Hames, B. D. (1981) An introduction to polyacrylamide gel electrophoresis, in Gel Electrophoresis of Proteins: A Practical Approach (Hames, B. D. and Rickwood, D. J., eds.) IRL, Oxford, pp. 1–91.
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© 1992 The Humana Press, Inc., Totowa, NJ
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Forster, S.J., Carr, F.J., Harris, W.J., Hamilton, A.A. (1992). Expression of Foreign Genes in Mammalian Cells Using an Antibody Fusion System. In: Manson, M.M. (eds) Immunochemical Protocols. Methods in Molecular Biology, vol 10. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-204-3:461
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DOI: https://doi.org/10.1385/0-89603-204-3:461
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-204-0
Online ISBN: 978-1-59259-497-9
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