A practical method for rescuing desired hybridomas during monoclonal antibody production |
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Authors: | Grazyna Adamus Z Suzanne Zam Scotts Emerson Paul A Hargrave |
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Institution: | (1) Department of Ophthalmology, University of Florida College of Medicine, Box J-284, 32610 Gainesville, FL;(2) Department of Biochemistry and Molecular Biology, University of Florida, 32610 Gainesville, Florida;(3) Department of Statistics, University of Florida, 32610 Gainesville, Florida |
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Abstract: | 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.
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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Keywords: | monoclonal antibody hybridoma limiting dilution cloning |
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