Publication: Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma
Issued Date
2002-09-12
Resource Type
ISSN
14726750
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2-s2.0-0037808989
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Mahidol University
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SCOPUS
Bibliographic Citation
BMC Biotechnology. Vol.2, (2002)
Suggested Citation
Duanpen Sandee, Sumalee Tungpradabkul, Manae Tsukio, Tadayuki Imanaka, Masahiro Takagi Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma. BMC Biotechnology. Vol.2, (2002). doi:10.1186/1472-6750-2-16 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/20359
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Title
Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma
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 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. © 2002 Sandee et al; licensee BioMed Central Ltd.