Selection and characterization of human antibodies against hepatitis B virus surface antigen (HBsAg) by phage-display.
Hepatitis B virus (HBV) is one of the main pathogens of hepatitis and hepatocarcinoma. Human plasma-derived antibody to HBV is being used as a prophylactic for postexposure to HBV and liver transplantation currently. However, it is required to replace the plasma-derived anti-HBs antibody (Ab) to a recombinant antibody because of limited availability of human plasma with high anti-HBs Ab titer and possible contamination of human pathogens. We constructed an anti-HBs Ab-enriched phage-display library from peripheral blood B cells of vaccinated volunteers and the size of library was approximately 1.0 x 10(7). The library was panned against hepatitis B surface antigen (HBsAg) and five different clones were isolated. All five clones exhibited the same heavy chain sequence; in contrast, light-chain exhibited one lambda and four different kappa sequences. The Fabs were expressed soluble in E. coli and exhibited affinities of 2.1 x 10(8) approximately 7.7 x 10(8) M(-1).
Kim
Se-Ho
SH
Central Research Center of the Korea Green Cross Corp, Kyunggi-Do, 449-903, Korea. sehokim@greencross.com
Park
Song-Yong
SY
D001483:Base Sequence; D001667:Binding, Competitive; D017931:DNA Primers / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007128:Immunoglobulin Fragments; D017912:Inovirus; D007700:Kinetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D019151:Peptide Library; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D006514:Hepatitis B Surface Antigens; D007128:Immunoglobulin Fragments; D019151:Peptide Library