Enhanced production of antibodies utilizing insolubilized immune complexes
First Claim
1. A method for producing a monoclonal antibody specific for an epitope distinct from that recognized by an existing monoclonal antibody, comprising the steps of:
- absorbing a first monoclonal antibody directed against a first epitope of a polyvalent antigen onto an inert, insoluble matrix capable of binding immunoglobulin, thereby forming an immunosorbent;
combining said immunosorbent with said polyvalent antigen, forming an insolubilized immune complex wherein said first epitope is masked by said first monoclonal antibody;
immunizing an animal with said insolubilized immune complex;
fusing spleen cells from said immunized animal to myeloma cells to form a hybridoma capable of producing a second monoclonal antibody directed against a second epitope of said polyvalent antigen;
culturing said hybridoma to produce said second monoclonal antibody; and
collecting said second monoclonal antibody as a product of said hybridoma.
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Abstract
A method for enhancing production of antibodies through immunization with insolubilized immune complexes is disclosed. Purified antigen or heterogeneous antigen mixtures may be combined with polyclonal or monoclonal antibody and the resultant complex bound to an insolubilized matrix to form insolubilized immune complexes. Methods for improving the immunogenicity of a soluble antigen and for producing monoclonal anti-iodiotypic antibodies are also disclosed. Monoclonal antibodies that are specific for a distinct, as yet unrecognized epitope may be produced by another disclosed method . Insolubilized immune complexes, comprising antigen and antibody that is ether directly linked to Sepharose® or absorbed onto insolubilized protein A, and immunosorbents, comprising antibody absorbed onto insolubilized protein A, are also disclosed.
15 Citations
16 Claims
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1. A method for producing a monoclonal antibody specific for an epitope distinct from that recognized by an existing monoclonal antibody, comprising the steps of:
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absorbing a first monoclonal antibody directed against a first epitope of a polyvalent antigen onto an inert, insoluble matrix capable of binding immunoglobulin, thereby forming an immunosorbent; combining said immunosorbent with said polyvalent antigen, forming an insolubilized immune complex wherein said first epitope is masked by said first monoclonal antibody; immunizing an animal with said insolubilized immune complex; fusing spleen cells from said immunized animal to myeloma cells to form a hybridoma capable of producing a second monoclonal antibody directed against a second epitope of said polyvalent antigen; culturing said hybridoma to produce said second monoclonal antibody; and collecting said second monoclonal antibody as a product of said hybridoma. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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- 11. The cell line NR-LU-01 A5791.
- 13. The cell line NR-LU-02 A5792.
- 15. The cell line NR-LU-03 A5793.
Specification