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Screen for human single chain variable region in antibody against human hepatitis C virus core protein binding protein 6
Yan-Wei Zhong, Jun Cheng, Zhong-Dong Zhang, Min Sun, Qiang Li, Ke Li, Lin Wang, Li Li, Ling-Xia Zhang, Ju-Mei Chen
Yan-Wei Zhong, Jun Cheng, Zhong-Dong Zhang, Min Sun, Qiang Li, Ke Li, Lin Wang, Li Li, Ling-Xia Zhang, Ju-Mei Chen, Gene Therapy Research Center, Institute of Infectious Diseases, The 302 Hospital of PLA, Beijing 100039, China
Supported by: Grants from National Natural Scientific Foundation of China, No. C639970674; No. C03011402.
Correspondence to: Jun Cheng, MD, PhD, Professor, Gene Therapy Research Center, Institute of Infectious Diseases, The 302 Hospital of PLA, Beijing 100039, China. cj@genetherapy.com.cn
Received: October 29, 2002 Revised: November 17, 2002 Accepted: November 28, 2002 Published online: April 15, 2003
AIM
To screen human single chain variable fragment (scFv) in antibody against hepatitis C virus (HCV) core protein-binding protein 6 (HCBP6) by phage display technique to determine HCBP6 in human liver tissue.
METHODS
The semisynthetic phage antibody library was panned by HCBP6 peptide which was coated on a microtiter plate, after five rounds of bio-panning, 48 clones specific for HCBP6 were determined with the enzyme-linked immunosorbent assay (ELISA). The specificity of HCBP6 scFv was identified by ELISA, cross reaction with bovine serum albumin (BSA)and competition inhibition assay. The DNA sequence of the positive clone was determined.
RESULTS
Specific phage antibody against HCBP6 was enriched by 162 times after five rounds of panning with HCBP6 peptide. Binding activities of 48 clones with HCBP6 peptide were determined by ELISA, and the results indicated that 30 clones could bind to HCBP6 peptide, 7 clones had low absorbance value at A450 nm; Direct and competition inhibition ELISA showed that one clone named P24, exhibited specific binding to HCBP6 peptide. Data of DNA sequence showed that the scFv gene encoded 771 bp.
CONCLUSION
The scFv fragments to HCBP6 can be successfully selected by phage display technique, which paves a way for study of distribution of HCBP6 in human liver tissues.
Key Words: N/A
Citation: Zhong YW, Cheng J, Zhang ZD, Sun M, Li Q, Li K, Wang L, Li L, Zhang LX, Chen JM. Screen for human single chain variable region in antibody against human hepatitis C virus core protein binding protein 6. Shijie Huaren Xiaohua Zazhi 2003; 11(4): 389-393
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