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US 8,999,707 B2Granted
Method of making hybrid cells that express useful antibodies
AssigneeThomas Jefferson University
InventorsScott K. Dessain, Sharad P. Adekar
Life of this patent
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Abstract
This invention relates to a novel hybridoma strategy that uses CD27+ B cells cultured in vitro to induce IgM to IgG class switch prior to fusion with a fusion partner. Hybridomas resulting from the fusion between CD27+ B cells and a fusion partner cell line and antibodies secreted from the hybridomas are included in the invention.
Claims13
A method of making a hybridoma, said method comprising
- culturing peripheral blood mononuclear cells (PBMCs) enriched for CD27+ PBMCs in the presence of CD40L and cytokines comprising IL-4 and IL-10 for a period of time in vitro,
- combining said cultured PBMCs enriched for CD27+ PBMCs with fusion partner cells, and
- fusing said cultured PBMCs enriched for CD27+ PBMCs with said fusion partner cells, whereby fusion of CD27+ B cells of said cultured PBMCs with said fusion partner cells produces monoclonal antibody-producing hybridomas.
A method of producing a monoclonal antibody, the method comprising:
- culturing peripheral blood mononuclear cells (PBMCs) enriched for CD27+ PBMCs in the presence of CD40L and cytokines comprising IL-4 and IL-10 for a period of time in vitro,
- combining said cultured PBMCs enriched for CD27+ PBMCs with fusion partner cells,
- fusing CD27+ B cells of said PBMCs enriched for CD27+ PBMCs with said fusion partner cells,
- selecting a hybridoma that produces said monoclonal antibody, and
- culturing said hybridoma to produce said monoclonal antibody.
2 more elements
A method of making a library of hybridomas, said method comprising
- culturing PBMCs enriched for CD27+ PBMCs in the presence CD40L and cytokines comprising IL-4 and IL-10 for a period of time in vitro,
- combining said cultured PBMCs enriched for CD27+ PBMCs with fusion partner cells, and
- fusing said cultured PBMCs enriched for CD27+ PBMCs with said fusion partner cells, whereby fusion of CD27+ B cells of said cultured PBMCs with said fusion partner cells produces a library of monoclonal antibody-producing hybridomas from said CD27+ B cells and said fusion partner cells.
Dependencies are inferred from the claim text.