Steven Foung

Publication Details

  • DEVELOPMENT OF MICROFUSION TECHNIQUES TO GENERATE HUMAN HYBRIDOMAS JOURNAL OF IMMUNOLOGICAL METHODS Foung, S., Perkins, S., Kafadar, K., Gessner, P., Zimmermann, U. 1990; 134 (1): 35-42

    Abstract:

    The rarity of antigen-specific B cells in peripheral blood and lymphoid tissues is a major limitation in the production of human monoclonal antibodies. This has led to a requirement for techniques capable of fusing small numbers of cells and achieving a higher hybridoma formation efficiency than currently is possible. The approach used in these studies to generate human hybridomas is based on the observation that under hypo-osmolar conditions electric field induced cell fusion or electrofusion is facilitated. Electrofusion parameters have been defined in strongly hypo-osmolar solutions which have resulted in a hybridoma formation efficiency greater than 5 X 10(-3) under optimal conditions. Furthermore, this has been accomplished with total input B cells of 1-2 X 10(5). This is a ten-fold reduction in the required number of input B cells and is associated with a hybridoma formation efficiency at least equal to that achieved with a higher input B cell number. An important factor in the development of this microfusion technique appears to be the duration of exposure to the hypo-osmolar solution by B cells to be immortalized. Other parameters which may affect hybridoma yield include the electrical field strength used for cell alignment and membrane breakdown, ratio of human B cells to fusion partner, washing procedure, post-fusion incubation time, and the elimination of toxic molecules.

    View details for Web of Science ID A1990EG52200003

    View details for PubMedID 2172386

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