关键词: 肝炎病毒, 乙型, 肝炎表面抗原, 乙型, 细菌噬菌体, 抗体库

Abstract: Objective To obtain the completely humanized single-chain antibody(scFv) against PreS1 of hepatitis B virus(HBV). Methods A 10¹⁰ cfu naive combinatorial library of phage-display human scFv, genes of which were derived from peripheral blood lymphocytes(PBLs) of 60 healthy donors, was constructed and panned by PreS1 of HBV. After 3 rounds of panning, the activity of individual clone was examined by ELISA. The clone with the highest A₄₁₀ value was expressed in E. coli HB2151 then identified by Western blotting. Sequencing was also carried out. Results After 3 rounds of panning, a 100 folds increase in the phage yielding relative to the first panning was noted. Measurement of antibody-antigen reaction revealed that scFv mono-clone specific to PreS1 with A₄₁₀=1.0 was enriched, which proportion of insert was verified by Western after being expressed. The sequencing result demonstrated further that it was a human scFv gene with which V_H belonged to human V_H1 family, and V_λ belonged to V_λ1. Conclusion The technique described in this paper is a powerful way to obtain humanized scFv gene against PreS1 of HBV, which may provide a more satisfactory therapy for hepatitis B.

Key words: hepatitis B virus, hepatitis B surface antigens, bacteriophages, antibody library