AI Article Synopsis
- The study describes the creation of hybridomas (cells that produce antibodies) from a small number of specific B cells that recognize HIV, using techniques like panning and immunobead selection.
- Researchers achieved a recovery rate of 5% and 24% purity through panning, and a better recovery of 17% but lower purity of 7% with immunobeads.
- The optimized process allows for the generation of up to 78 hybridomas from a single B cell, with a successful B cell immortalization efficiency of up to 10%.
Article Abstract
This paper reports the generation of monoclonal antibody producing hybridomas from a small number of antigen-specific B cells selected by panning on antigen-coated dishes and rosetting with antigen-coupled paramagnetic beads. Anti-HIV positive B cells from spleen could be recovered by panning with an efficiency of 5% and a purity of 24%. Immunobead selection of anti-HIV positive B cells from the same mice yielded a recovery of 17% and a purity of 7%. Various experimental conditions with respect to the selection of specific B cells were investigated, leading to an optimized protocol for the isolation of a limited subset of B cells. The selected cells retained their property to produce immunoglobulins and could be clonally expanded in the presence of human T cell supernatant and irradiated murine thymoma helper cells to generate sufficient cells for a mini-electrofusion with NS-1 myeloma cells. Up to 78 specific hybridomas could be generated from one anti-HIV positive B cell. An overall efficiency of specific B cell immortalization of up to 10% was obtained.
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| http://dx.doi.org/10.1016/0022-1759(93)90236-z | DOI Listing |
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