Selection and characterization of human antibodies against hepatitis B virus surface antigen (HBsAg) by phage-display
Selection and characterization of human antibodies against hepatitis B virus surface antigen (HBsAg) by phage-display
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DOI：
10.1089/153685902761022742
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发表时间：
2002-10-01
期刊：
影响因子：
--
通讯作者：
Park, SY
中科院分区：
文献类型：
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作者：
Kim, SH;Park, SY
乙型肝炎病毒（HBV）是导致肝炎和肝癌的主要病原体之一。目前，人血浆源性乙肝病毒抗体被用作乙肝病毒暴露后和肝移植的预防药物。然而，由于具有高抗hbs抗体滴度的人血浆有限，且可能污染人病原体，因此需要用重组抗体替代血浆源性抗hbs抗体（Ab）。我们利用接种疫苗的志愿者外周血B细胞构建了一个抗hbs抗体富集的噬菌体展示文库，文库大小约为1.0 × 10(7)。对该文库进行乙型肝炎表面抗原（HBsAg）筛选，分离出5个不同的克隆。5个克隆均表现出相同的重链序列；而轻链则表现出1个lambda和4个不同的kappa序列。这些fab可在大肠杆菌中表达，亲和性为2.1 × 10(8)，与7.7 × 10(8) M-1相似。
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 similar to1.0 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) similar to 7.7 x 10(8) M-1.