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Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma

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Author(s): Sandee Duanpen | Tungpradabkul Sumalee | Tsukio Manae | Imanaka Tadayuki | Takagi Masahiro

Journal: BMC Biotechnology
ISSN 1472-6750

Volume: 2;
Issue: 1;
Start page: 16;
Date: 2002;
Original page

Keywords: Single-chain Fv (scFv) | Hepatocellular carcinoma (HCC) | Bacterial expression | Inclusion body

ABSTRACT
Abstract Background Hep27 monoclonal (Hep27 Mab) is an antibody against hepatocellular carcinoma. Hep27 Mab itself can inhibit the growth of a hepatocellular carcinoma cell line (HCC-S102). We attempted to produce a single-chain fragment (scFv), a small fragment containing an antigen-binding site of Hep27 Mab, by using DNA-recombinant techniques. Results The sequences encoding the variable regions of heavy (VH) and light (VL) chains of a murine Hep27 Mab were linked together by a linker peptide (Gly4Ser)3 and tagged with a hexa-histidine at the C-terminal; the resultant DNA construct was expressed in E. coli as an insoluble protein. The denatured scFv was refolded and purified by immobilized metal ion affinity chromatography (12 mg/l with a molecular weight of 27 kDa). Hep27scFv exhibited a tumoricidal activity against the HCC-S102 cell as its parental antibody (Hep27 Mab). Conclusion This scFv may be a potential candidate for a targeting agent in HCC immunodiagnosis or immunotherapy.
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