Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma
<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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2002-09-01
|
| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | http://www.biomedcentral.com/1472-6750/2/16 |
| Summary: | <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<sub>H</sub>) and light (V<sub>L</sub>) chains of a murine Hep27 Mab were linked together by a linker peptide (Gly4Ser)<sub>3</sub> 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> |
|---|---|
| ISSN: | 1472-6750 |