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Functional Expression of Human Monoclonal Antibody Genes Directed Against Pseudomonal Exotoxin A in Mouse Myeloma Cells

Bio/Technology volume 7, pages 805–810 (1989)Cite this article

Abstract

We have cloned immunoglobulin (Ig) genes from an Epstein-Barr (EB) virus-transformed human B cell line, FK-001, which produced a monoclonal human IgM that specifically binds Pseudomonas aeruginosa exotoxin A (Ex-A). Cloned Ig genomic DNA was introduced into Ig non-producing mouse myeloma cells, P3-X63-Ag-8-6.5.3. (653) and Sp2/0-Ag14 (Sp2/0), by protoplast fusion or electroporation using a series of pSV2 vectors. Transfectant-derived recombinant antibodies (r-Abs) and antibodies produced by the original cell FK-001 did not differ in binding specificity or affinity to Ex-A. Recombinant antibodies and those of FK-001 were pentameric although joining (J) genes of human origin were not introduced into the mouse myeloma cells. Heterogeneity in the molecular weights of r-Abs expressed in different mouse myeloma cells was observed, and shown to be due to differential N-glycosylation of the μ-chain polypeptides. Thus, human Ig μ and κ genomic DNA with its own promoters and enhancers can function efficiently in mouse myeloma cells in vivo resulting in transfectants that synthesize active human IgM.

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Author notes

  1. Hiroshi Noguchi: Corresponding author.

Authors and Affiliations

  1. Sumitomo Chemical Co., Ltd., Takarazuka Research Center, Laboratory of Biotechnology, 4-2-1, Takatsukasa, Takarazuka, Hyogo, 665, Japan

    Tomoyuki Nakatani, Noriko Nomura, Kazuhiko Horigome, Hiroshi Ohtsuka & Hiroshi Noguchi

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  1. Tomoyuki Nakatani
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  2. Noriko Nomura
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  3. Kazuhiko Horigome
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  4. Hiroshi Ohtsuka
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  5. Hiroshi Noguchi
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Nakatani, T., Nomura, N., Horigome, K. et al. Functional Expression of Human Monoclonal Antibody Genes Directed Against Pseudomonal Exotoxin A in Mouse Myeloma Cells. Nat Biotechnol 7, 805–810 (1989). https://doi.org/10.1038/nbt0889-805

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