Expression of an antibody-enzyme complex by the L-chain fusion method(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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In this report, we describe a novel method for directly preparing enzyme-labeled antibodies harvested from IgM-producing hybridoma cells. We constructed expression vectors for antibody light (L) chain-enzyme fusion proteins by linking either the genes for the murine lambda L chain or its constant region (C_L) with one of two proteins, either the secreted placental alkaline phosphatase or Gaussia luciferase (Gluc). When the vectors were transfected into anti-NP (4-hydroxy-3-nitrophacetyl) IgM-producing myeloma cells, secretion of the IgM-enzyme complex from the gene-transfected cells was confirmed by a direct enzyme-linked immunosorbent assay with an immobilized antigen. Furthermore, when human hybridoma HF10B4, a cell line that produces anti-human lung cancer IgM, was transfected with the vector containing L-Gluc, a significantly stronger signal was obtained for the human lung carcinoma SBC-1 cells than for cervical HeLa cells. Because successful production of an active IgM-enzyme complex containing a heterologous L chain-enzyme fusion was observed, the L-chain fusion method will be a generally applicable method for preparing various IgM-enzyme complexes.
- 2013-07-00
著者
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Ueda Hiroshi
Department Of Anatomy School Of Medicine University Of Occupational And Environmental Health
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Ueda Hiroshi
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo:Department of Bioengineering, School of Engineering, The University of Tokyo:(Present office)Chemical Resources Laboratory, Tokyo Institute of Technology
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Han Chungyong
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo
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Ihara Masaki
Department of Bioengineering, School of Engineering, The University of Tokyo
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Ihara Masaki
Department of Bioengineering, School of Engineering, The University of Tokyo:(Present office)Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University
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