Duanpen SandeeSumalee TungpradabkulManae TsukioTadayuki ImanakaMasahiro TakagiOsaka UniversityMahidol UniversityKyoto UniversityJapan Advanced Institute of Science and Technology2018-07-242018-07-242002-09-12BMC Biotechnology. Vol.2, (2002)147267502-s2.0-0037808989https://repository.li.mahidol.ac.th/handle/20.500.14594/20359Background: 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.Mahidol UniversityMedicineArticleSCOPUS10.1186/1472-6750-2-16