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A practical method for rescuing desired hybridomas during monoclonal antibody production

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Summary

We present a practical method for the rescue of previosly stable hybridoma clones which increases the proportion of desired cells in the population before cloning by limiting dilution. When the antibody activity of a culture supernatant was lower than that previously obtained, a precloning distribution at a density of 10 cells per microtiter well greatly improved the chances of obtaining a single active clone by subsequent limiting dilution. The Poisson distribution model was used to evaluate the method. Probabilities calculated clearly demonstrate the advantage of this precloning distribution step when attempting to isolate a hydridoma cell line that is relatively rare in a population.

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Authors and Affiliations

  1. Department of Ophthalmology, University of Florida College of Medicine, Box J-284, 32610, Gainesville, FL

    Grazyna Adamus, Z. Suzanne Zam & Paul A. Hargrave

  2. Department of Biochemistry and Molecular Biology, University of Florida, 32610, Gainesville, Florida

    Paul A. Hargrave

  3. Department of Statistics, University of Florida, 32610, Gainesville, Florida

    Scotts Emerson

Authors
  1. Grazyna Adamus
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  2. Z. Suzanne Zam
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  3. Scotts Emerson
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  4. Paul A. Hargrave
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Additional information

This work was supported in part by grants EY 06225 and EY 06226 from the National Eye Institute of the National Institutes of Health, Bethesda, MD and by an unrestricted departmental award from Research to Prevent Blindness, Inc.

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Adamus, G., Zam, Z.S., Emerson, S. et al. A practical method for rescuing desired hybridomas during monoclonal antibody production. In Vitro Cell Dev Biol 25, 1141–1146 (1989). https://doi.org/10.1007/BF02621265

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  • DOI: https://doi.org/10.1007/BF02621265

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