Bacterial cell surface display for epitope mapping of hepatitis C virus core antigen.
Cell surface expression of protein has been widely used to display enzymes and antigens. Here we show that Pseudomonas syringae ice nucleation protein with a deletion of internal repeating domain (INC) can be used in Escherichia coli to display peptide in a conformationally active form on the outside of the folded protein by fusing to the C-terminus of INC. Diagnostic potential of this technology was demonstrated by effective mapping of antigenic epitopes derived from hepatitis C virus (HCV) core protein. Amino acids 1-38 and 26-53 of HCV core protein were found to react more sensitively in a native conformation with the HCV patient sera than commercial diagnostic antigen, c22p (amino acids 10-53) by display-ELISA. These results demonstrate that the bacterial cell surface display using INC is useful for peptide presentation and thus epitope mapping of antigen.
Kang
Su-Min
SM
Department of Biotechnology, Yonsei University, 134 Sinchon-dong, Seodaemun-gu, Seoul 120-749, South Korea.
Rhee
Jin-Kyu
JK
Kim
Eui-Joong
EJ
Han
Kwang-Hyub
KH
Oh
Jong-Won
JW
D000954:Antigens, Surface / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D005798:Genes, Bacterial; D005814:Genes, Viral; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000175:diagnosis / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood / Q000276:immunology; D018936:Hepatitis C Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008856:Microscopy, Fluorescence; D044224:Pseudomonas syringae / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D014169:Transformation, Bacterial / Q000235:genetics; D014758:Viral Core Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C063113:ice nucleation protein; C069130:nucleocapsid protein, Hepatitis C virus