A novel recombinant multiepitope protein as a hepatitis C diagnostic intermediate of high sensitivity and specificity

Protein Expression and Purification
Chugh A DiptiKhanna Navin

Abstract

A novel recombinant multiepitope protein has been designed that consists of six linear, immunodominant, and phylogenetically conserved epitopes from hepatitis C virus. Five of these antigens (core, NS3, NS4I, NS4II, and NS5) are being used in many of the third-generation kits while sixth epitope (core3g) is an additional sequence from a newly identified Indian isolate. The genes for these epitopes have been joined together to code for a single multiepitope protein that has been evaluated for its diagnostic potential for the detection of anti-HCV antibodies in human plasma. Two separate synthetic genes have been designed, both encoding the same six epitopes in a single open reading frame along with spacers having additional amino acids to function as flexible (r-HCV-F-MEP) or rigid (r-HCV-R-MEP) linkers. High-level expression of hepatitis C multiepitope protein in Escherichia coli has been achieved. The protein has been purified using a single affinity step yielding >25 mg pure protein/liter culture and used as the coating antigen in anti-HCV EIA. The use of this multiepitope protein eliminates the requirement for multiple diagnostic intermediates for the development of anti-HCV diagnostic kit. The sensitivity and specificity of...Continue Reading

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Citations

Mar 7, 2014·Applied Microbiology and Biotechnology·Xue Jun WangRong Yue Cao
Mar 5, 2016·Hepatitis Research and Treatment·Alexsandro S GaldinoFernando A G Torres
Dec 1, 2015·Journal of Virological Methods·Teppo SalminenSheikh M Talha
Jan 15, 2011·Journal of Virological Methods·Sheikh M TalhaNavin Khanna
Apr 17, 2013·Biotechnology and Applied Biochemistry·Chandrasekhar GurramkondaKrothapalli Raja Surya Sambasiva Rao
Mar 1, 2014·Biotechnology and Applied Biochemistry·Daylen Aguilar-NoriegaSantiago Dueñas-Carrera
Mar 19, 2015·Journal of Medical Virology·Reza TaherkhaniManoochehr Makvandi
Dec 3, 2013·BioMed Research International·Marilen Queiroz de SouzaMaria Sueli Soares Felipe
Jan 9, 2015·PLoS Neglected Tropical Diseases·Angélica Rosa FariaHélida M Andrade
Feb 18, 2018·Journal of Clinical Laboratory Analysis·Ronaldo Luis ThomasiniMauro Martins Teixeira
Jul 25, 2019·Recent Patents on Biotechnology·Patrícia A F RibeiroAlexsandro S Galdino
Jan 22, 2010·Clinical and Vaccine Immunology : CVI·Sheikh M TalhaNavin Khanna
Jun 1, 2018·Current Medicinal Chemistry·Inese MierinaMaris Turks

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