Human-murine hybridoma fusion partner
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsEdgar G. Engleman, Steven K. H. Foung, F. Carl Grumet
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Abstract
By careful screening and mutation, a human-murine hybridoma suitable as a fusion partner for immortalizing an antibody-secreting B cell has been generated. The trioma fusion products of this immortalizing partner are stable producers of human monoclonal antibodies. A trioma which produces monoclonal human anti-varicella zoster is disclosed.
Claims14
An immortalizing fusion partner for use in producing a trioma cell line capable of secreting a human monoclonal antibody specific against a selected antigen, when fused with a non-malignant b-lymphoid cell derived from a human donor exposed to such antigen, comprising a fused mouse myeloma/non-malignant human B-lymphocyte cell line which (a) expresses HLA surface antigens, (b) does not secrete immunoglobulins, and (c) is deficient in hypoxanthine phosphoribosyl transferase, as evidenced by the inability of the cell line to grow in hypoxanthine-aminopterin-thymidine or azaserine-hypoxanthine medium.
A trioma cell line capable of secreting a normal human monoclonal antibody specific against a selected antigen, said cell line comprising the fusion product of:
- (1) a mouse myeloma/non-malignant human B-lymphocyte hybridoma fusion partner which (a) expresses HLA surface antigens, (b) does not secrete immunoglobulins, and (c) is deficient in hypoxanthine phosphoribosyl transferase, as evidenced by the inability of the fusion partner to grow in hypoxanthine-aminopterin-thymidine or azaserine-hypoxanthine medium, and
- (2) a non-malignant B-lymphoid cell derived from a human donor exposed to the selected antigen.
A method of producing a human monoclonal antibody which is specific against a selected antigen, comprising
- providing a fusion partner produced by fusing mouse myeloma cells and non-malignant human B-lymphocytes, selecting fusion products which show stable immunoglobulin secretion and HLA surface antigen production in culture, treating the selected fusion products with a mutagen, and selecting mutagenized fusion products which retain the ability to produce HLA surface antigen, show no immunoglobulin secretion, and are unable to survive in a growth medium which allows growth of a successful fusion product formed by fusing the fusion partner with a human immunoglobulin-secreting B-lymphoid cell,
- fusing the fusion partner with a non-malignant B-lymphoid cell derived from a human donor exposed to the selected antigen, and
- selecting fusion products which are viable in such medium and which secrete antigen-specific monoclonal antibody.
The method of claim:
- 12, wherein the antibody is made by a fusion product of the fusion partner and the B-lymphoid cells having the characteristics of ATCC HB 8463.
Dependencies are inferred from the claim text.