title: Hepatitis B virus-neutralizing anti-pre-S1 human antibody fragments from large naïve antibody phage library.
abstract: We report the construction of a large nonimmunized human phage antibody library in single-chain variable region fragment (scFv) format, which allowed the selection of antibodies that neutralize hepatitis B virus (HBV) in vitro. We generated 1.1 x 10(10) independent scFv clones using the cDNA of functional variable (V) gene segments of heavy and light chains purified from the peripheral blood mononuclear cells of 50 nonimmunized human donors. Using BIAcore, we selected two clones that recognized pre-S1 and neutralized pre-S1 and HBV binding to Chang liver cells. Clone G10 had the highest affinity (K(D)=1.69 x 10(-7)M), which was higher than that of clone 1E4 that was generated previously from a heavy chain-shuffled immune library. The off-rates of clones were within 10(-3)s(-1) as determined by BIAcore and were comparable to those of antibodies derived from a normal secondary immune response. In the inhibition assays of pre-S1 and virus binding to Chang liver cells using flow cytometry and the polymerase chain reaction, G10 had better neutralizing activity than 1E4. The new phage library may be a valuable source of antibodies with reasonable affinities to different targets, and the anti-pre-S1 G10 may be a good candidate for immunoprophylaxis against HBV infection.
authors.affiliation: Department of Microbiology, College of Medicine and Center for Viral Disease Research, Inje University, Jin-Gu, Busan 614-735, Republic of Korea.
authors.forename: Sae-Gwang
authors.initials: SG
authors.lastname: Park
authors.forename: Yong-Joo
authors.initials: YJ
authors.lastname: Jeong
authors.forename: Yong-Yi
authors.initials: YY
authors.lastname: Lee
authors.forename: Ik-Jung
authors.initials: IJ
authors.lastname: Kim
authors.forename: Su-Kil
authors.initials: SK
authors.lastname: Seo
authors.forename: Eui-Joong
authors.initials: EJ
authors.lastname: Kim
authors.forename: Heung-Chae
authors.initials: HC
authors.lastname: Jung
authors.forename: Jae-Gu
authors.initials: JG
authors.lastname: Pan
authors.forename: Sung-Jae
authors.initials: SJ
authors.lastname: Park
authors.forename: Yeon-Jae
authors.initials: YJ
authors.lastname: Lee
authors.forename: Ik-Sang
authors.initials: IS
authors.lastname: Kim
authors.forename: In-Hak
authors.initials: IH
authors.lastname: Choi
mesh_terms: D001435:Bacteriophages; D002460:Cell Line; D006510:Hepatitis B Antibodies / Q000096:biosynthesis* / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics*; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D019151:Peptide Library; D011498:Protein Precursors / Q000276:immunology*
chemical_list: D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011498:Protein Precursors; C050806:presurface protein 1, hepatitis B surface antigen