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 hexa-histidine 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.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC128817 | PMC |
| http://dx.doi.org/10.1186/1472-6750-2-16 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Tumor suppressive monoclonal antibody belonging to the V(H) 7183 family directed to the oncodevelopmental carbohydrate antigen on human hepatocellular carcinoma.
J Biosci Bioeng
November 2005
Department of Biotechnology, Graduate school of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
Hep27 monoclonal antibody (Hep27 Mab) was raised by immunizing BALB/c mice with cells of the Thai human hepatocellular carcinoma (HCC) cell line HCC-S102 using hybridoma technology. The Hep27 Mab recognizes oncofetal development antigens by reacting with many HCC, other cancers, fetal and newborn liver but not adult liver. The Hep27 Mab alone markedly inhibits the growth of hepatocellular carcinoma cell lines (65% viability on the third day), suggesting its clinical usefulness.
View Article and Find Full Text PDFCombination of Dsb coexpression and an addition of sorbitol markedly enhanced soluble expression of single-chain Fv in Escherichia coli.
Biotechnol Bioeng
August 2005
Department of Biotechnology, Graduate school of Engineering, Osaka University, Suita, Osaka, Japan.
Many eukaryotic proteins have been produced successfully in Escherichia coli. However, not every gene can be expressed efficiently in this organism. Most proteins, especially those with multiple disulfide bonds, have been shown to form insoluble protein or inclusion body in E.
View Article and Find Full Text PDFConstruction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma.
BMC Biotechnol
September 2002
Department of Biotechnology, Graduate school of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
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.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!