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Detection and Selection of Cultured Cells Secreting Recombinant Product by Soft Agar Cloning and Antibody Overlay

  • Protocol
Recombinant Protein Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 63))

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Abstract

Detection of expression of recombinant genes in transfected cells can be combined with cell cloning to achieve pure cell populations secreting high levels of heterologous protein. The combination of cloning and selection speeds up the generation of permanent cell lines, increases the number of potential insertion events that can be screened, and reduces the tedium of sequentially cloning then analyzing the production levels of each clone.

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References

  1. Gherardi, E., Pannell, R., and Milstein, C. (1990) A single-step procedure for cloning and selection of antibody-secreting hybridomas. J. Immunol. Meth. 126, 61–68.

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  2. Walls, J. D. and Grinnell, B. W. (1990) A rapid and versatile method for the detection and isolation of mammalian cell lines secreting recombinant proteins. BioTechniques 8, 138–142.

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  3. Gibson, M. G., Ashey, L., Newell, A. H., and Bradley, C. (1993) Soft agar cloning with antibody overlay to identify chinese hamster ovary clones secreting recombinant products. BioTechniques 15, 594–597.

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  4. Coffino, P. and Scharff, M. D. (1971) Rate of somatic mutation in immunoglobulin production by mouse myeloma cells. Proc. Natl. Acad. Sci. USA 68, 219–223.

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Author information

Authors and Affiliations

  1. Gene Therapy Program, University of California, San Diego, CA

    Marylou G. Gibson

  2. Immunex Corp., Seattle, WA

    Karmen Hodges & Lauretta Lowther

Authors
  1. Marylou G. Gibson
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  2. Karmen Hodges
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  3. Lauretta Lowther
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Editor information

Editors and Affiliations

  1. Thomas Jefferson University, Philadelphia, PA

    Rocky S. Tuan

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Copyright information

© 1997 Humana Press Inc., Totowa, NJ

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Gibson, M.G., Hodges, K., Lowther, L. (1997). Detection and Selection of Cultured Cells Secreting Recombinant Product by Soft Agar Cloning and Antibody Overlay. In: Tuan, R.S. (eds) Recombinant Protein Protocols. Methods in Molecular Biology™, vol 63. Humana Press. https://doi.org/10.1385/0-89603-481-X:77

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  • DOI: https://doi.org/10.1385/0-89603-481-X:77

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-481-5

  • Online ISBN: 978-1-59259-549-5

  • eBook Packages: Springer Protocols

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