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

• Main Authors: Imanaka Tadayuki, Tsukio Manae, Tungpradabkul Sumalee, Sandee Duanpen, Takagi Masahiro
• Format: Article
• Language: English
• Published: BMC 2002-09-01
• Series: BMC Biotechnology
• Subjects: Single-chain Fv (scFv); Hepatocellular carcinoma (HCC); Bacterial expression; Inclusion body
• Online Access: http://www.biomedcentral.com/1472-6750/2/16

<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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 hexa-histidine at the C-terminal; the resultant DNA construct was expressed in <it>E. coli</it> 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).</p> <p>Conclusion</p> <p>This scFv may be a potential candidate for a targeting agent in HCC immunodiagnosis or immunotherapy.</p>