Hepatitis B virus-neutralizing anti-pre-S1 human antibody fragments from large naïve antibody phage library.
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.
Park
Sae-Gwang
SG
Department of Microbiology, College of Medicine and Center for Viral Disease Research, Inje University, Jin-Gu, Busan 614-735, Republic of Korea.
Jeong
Yong-Joo
YJ
Lee
Yong-Yi
YY
Kim
Ik-Jung
IJ
Seo
Su-Kil
SK
Kim
Eui-Joong
EJ
Jung
Heung-Chae
HC
Pan
Jae-Gu
JG
Park
Sung-Jae
SJ
Lee
Yeon-Jae
YJ
Kim
Ik-Sang
IS
Choi
In-Hak
IH
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*
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