Generation of a recombinant mouse-human chimaeric monoclonal antibody directed against human carcinoembryonic antigen

Autor: Felix Businger, Franz Buchegger, L. Lee Gill, Jean-Pierre Mach, Ronald F.J. De Winter, Christoph Heusser, Kathrin Wagner, Melvyn Hollis, Domenico Ammaturo, Norman Hardman
Rok vydání: 1989
Předmět:
Zdroj: International Journal of Cancer. 44:424-433
ISSN: 1097-0215
0020-7136
DOI: 10.1002/ijc.2910440308
Popis: A procedure was devised for the identification and specific cloning of functionally rearranged variable region immunoglobulin (Ig) gene segments from genomic DNA of a murine hybridoma cell line which produces a high-affinity monoclonal antibody (MAb) directed against human carcinoembryonic antigen (CEA). The cloned, functionally-rearranged murine Ig H-chain and L-chain variable region gene segments were incorporated into plasmid vectors capable of directing the expression of a chimaeric mouse-human antibody molecule with human (gamma 4, kappa) constant region sequences. Expression plasmids were transfected into a mouse myeloma cell line by electroporation and transfectomas secreting functional chimaeric antibody selected. Chimaeric antibody generated by transfectomas was analysed and shown to compete effectively with its murine counterpart for binding to the CEA epitope, and to have an equivalent antigen-binding affinity. This anti-CEA recombinant antibody should find application in in vivo diagnosis by immunoscintigraphy of human colonic carcinoma, and possibly also in therapy of the disease, overcoming some of the difficulties associated with the repeated use of non-human immunoglobulins in human patients.
Databáze: OpenAIRE
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