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

• Sandee, Duanpen

• Tungpradabkul, Sumalee

• Tsukio, Manae

• Imanaka, Tadayuki

• Takagi, Masahiro

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 (V_H) and light (V_L) chains of a murine Hep27 Mab were linked together by a linker peptide (Gly4Ser)_3 and tagged with a hexahistidine 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.

identifier:https://dspace.jaist.ac.jp/dspace/handle/10119/8548

Journal

• BMC Biotechnology

  BMC Biotechnology 2 (article no.16), 2002-09-12

  BioMed Central Ltd.

Keywords

• Single-chain Fv (scFv)
• Hepatocellular carcinoma (HCC)
• Bacterial expression
• Inclusion body

Details

• CRID: 1050564287490736640
• NII Article ID: 120001746772
• ISSN: 14726750
• Web Site: http://hdl.handle.net/10119/8548
• Text Lang: en
• Article Type: journal article
• Data Source:
  • IRDB
  • CiNii Articles