Cloning, expression and identification by immunohistochemistry of humanized single-chain variable fragment antibody against hepatitis C virus core protein

• Autorzy: Lun Y. , Cheng J. , Zhong Y. , Yu Z. , Wang Q. , Wang F. , Feng J.
• Języki publikacji: EN

Abstrakty

EN

Expression of single-chain variable fragment (scFv) antibodies on the surface of bacteriophage is widely used to prepare antibodies with pre-defined specificities. A phage antibody library containing the gene for scFv antibody against Hepatitis C virus core protein was panned with core protein immobilized on microtiter plate wells. After five rounds of panning 60 phage clones specific to core protein were obtained and one selected clone was sequenced. It was found that the specifically detected antigen consists of 774bp and is capable of encoding 257 amino acids in the patients but not in healthy persons.

Słowa kluczowe

EN

hepatitis C virus; core protein; phage display; single-chain variable fragment antibody; immunohistochemistry; expression; identification; bacteriophage; antibody; cloning

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2011
• Tom: 60
• Numer: 1
• Opis fizyczny: p.13-17,fig.,ref.

Twórcy

autor

Lun Y.

• Liaoning Provincial University Key Laboratory of Biophysics, Medical College, Dalian University, Dalian 116622, China

autor

Cheng J.

autor

Zhong Y.

autor

Yu Z.

autor

Wang Q.

autor

Wang F.

autor

Feng J.

Bibliografia

• Aoyagi K., C. Ohue and K. Iida. 1999. Development of a simple and highly sensitive enzyme immunoassay for hepatitis C virus core antigen. J. Clin. Microbiol. 37: 1802­1808.
• Armstrong G.L. 2003. Commentary: modeling the epidemiology of hepatitis C and its complications. Int. J. Epidemiol. 32: 725­726.
• Canaan-Haden L., M. Ayala and M.E. Fernandez-de-Cossio. 1995. Purification and application of a single-chain Fv antibody fragment specific to hepatitis B virus surface antigen. Biotechniques 19: 606­612.
• Di Bisceglie A.M. 2000. Natural history of hepatitis C: its impact on clinical management. Hepatology 31: 1014­1018.
• Dickson R.C., M. Mizokami and E. Orito. 1999. Quantification of serum HCV core antigen by a fluorescent enzyme immunoassay in liver transplant recipients with recurrent hepatitis C: clinical and virologic implications. Transplantation. 68: 1512.
• Hoogenboom H.R., A.P. de Bruine and S.E. Hufton. 1998.
• Anti-body phage display technology and its applications. Immunotechnology 4: 1­20.
• Houghton M., A. Weiner and J. Han. 1991. Molecular biology of the hepatitis C virus: implications for diagnosis, development, and control of viral disease. Hepatology 14: 381.
• Kittlesen D.J., K.A. Chianese-Bullock and Z.Q. Yao. 2000.
• Interaction between complement receptor gC1qR and hepatitis C virus core protein inhibits T-lymphocyte proliferation. J. Clin. Investig. 106: 1239­1249.
• Hagedorn C.H. and C.M. Rice. 2000. The hepatitis C viruses. Springer-Verlag KG Berlin, Germany.
• Lamarre A. and P.J. Talbot. 1997. Characterization of phagedisplayed recombinant anti-idiotypic antibody fragments against coronavirus neutralizing monoclonal antibodies. Viral. Immunol. 10: 175-182.
• Large M.K., D.J. Kittlesen and Y.S. Hahn. 1999. Suppression of host immune response by the core protein of hepatitis C virus: possible implications for hepatitis C virus persistence. J. Immunol. 162: 931-938.
• Marks J.D., H.R. Hoogenboom and T.P. Bonnert. 1991. Byimmunization human antibodies from V-gene libraries displayed on phage. J. Mol. Biol. 222: 581-597.
• Menez R., M. Bossus and B.H. Muller. 2003. Crystal structure of a hydrophobic immunodominant antigenic site on hepatitis C virus core protein complexed to monoclonal antibody 19D9D6. J. Immunol. 170: 1917-1924.
• Nelson D.R., C.G. Marousis and G.L. Davis. 1997. The role of hepatitis C virus-specific cytotoxic lymphocytes in chronic hepatitis C. J. Immunol. 158: 1473-1481.
• Reed K.E. and C.M. Rice. 2000. Overview of hepatitis C virus genome stucture, polyprotein processing, and protein properties. Curr. Top. Microbiol. Immunol. 242: 55-84.
• Rondon U. and W.A. Marasco. 1997. Intracellular antibodies (intrabodies) for gene therapy of infectious disease. Annu. Rev. Microbiol. 51: 257-283.
• Songsivilai S., T. Dharakul and R. Kunkitti. 1996. Molecular cloning and expression of hepatitis C virus core protein and production of monoclonal antibodies to the recombinant protein. Asian Pac J Allergy Immunol. 14: 31-41.
• Stoll-Keller F., H. Barth and S. Fafi-Kremer. 2009. Development of hepatitis C virus vaccines: challenges and progress. Expert Rev. Vaccines 8: 333-345.
• Tanaka E., C. Ohue and K. Aoyagi. 2000. Evaluation of a new enzyme immunoassay for hepatitis C virus (HCV) ore antigen with clinical sensitivity approximating that of genomic amplification of HCV RNA. Hepatology 32: 388.
• Zhong Y., J. Cheng and S. Shi. 2001. Screening and characterization of human phage antibody to hepatitis C core antigen (in Chinese). Zhonghua Gan Zang Bing Za Zhi 9: 217-219.