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目的 :筛选能识别庚型肝炎病毒 (HGV) E2区的单克隆抗体和有竞争抑制活性的多肽。方法 :利用抗 HGV E2区的3株单克隆抗体 M6 ,M13,M30作为筛选配基 ,对构象限制性的随机环 9肽库进行亲和筛选 ,并对阳性噬菌体克隆进行功能鉴定。结果 :证实亲和筛选具有良好的富集效果后 ,随机挑出 16个噬菌体克隆进行交叉结合实验 ,有 14个克隆与 M6抗体有较强的特异结合 ;其中 10个克隆在竞争抑制实验中抑制率大于 5 0 %。 DNA测序表明 9个克隆具有氨基酸核心序列 GYAPL S,该序列与 HGV E2区第 30 6～ 311位氨基酸有较好的同源性。用合成肽与核心序列相似的噬菌体克隆 P9GC5和 P9GC11进行HGV E2抗原抗体实验 ,显示它们的 IC5 0 分别为 4.5和 7.2 nmol/ L。结论 :从噬菌体随机肽库中筛选到能与 HGV E2抗体特异性结合的 HGV抗原表位 ,并确定与合成了具有活性的核心多肽序列。
OBJECTIVE: To screen monoclonal antibodies recognizing the E2 region of Hepatitis G virus (HGV) and peptides with competitive inhibitory activity. Methods: Three monoclonal antibodies M6, M13 and M30 of anti-HGV E2 region were used as screening ligands, and the conformationally restricted random 9-peptide library was screened by affinity. The positive phage clones were identified. Results: After confirming that the affinity screening had a good enrichment effect, 16 phage clones were randomly selected for cross-binding experiments. Fourteen clones showed strong specific binding to M6 antibody. Among them, 10 clones were inhibited in competition inhibition experiments Rate is greater than 50%. DNA sequencing showed that nine clones had the amino acid core sequence GYAPL S, which had a good homology with the amino acids 306-611 of HGV E2. HGV E2 antigen antibody experiments with phage clones P9GC5 and P9GC11 with similar synthetic sequences to the core sequence showed that their IC50 were 4.5 and 7.2 nmol / L, respectively. Conclusion: The HGV antigen epitope that can specifically bind to HGV E2 antibody was screened from phage random peptide library, and the active core polypeptide sequence was identified and synthesized.