Discontinuous epitopes of hepatitis B surface antigen derived from a filamentous phage peptide library
Discontinuous epitopes of hepatitis B surface antigen derived from a filamentous phage peptide library
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DOI：
10.1073/pnas.93.5.1997
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发表时间：
1996-03-05
影响因子：
11.1
通讯作者：
Mandecki, W
中科院分区：
文献类型：
--
作者：
Chen, YCJ;Delbrook, K;Mandecki, W
用4株抗-HBs单抗的表位图谱研究了小分子乙肝病毒表面抗原(HBs)的结构。利用丝状噬菌体多肽文库，通过亲和浓缩实验(生物扫描)获得表位的氨基酸序列。该文库由10(9)个不同的克隆组成，含有一个与基因III融合的30个残基的多肽。通过对文库中的抗体进行淘洗获得的多肽与天然的乙肝表面抗原序列之间的序列同源性，使得结合区域的准确描述成为可能。四株单抗中有三株与乙肝表面抗原第101和207位氨基酸残基之间的不连续表位相结合。第四株mAb可与121-124位残基结合。序列数据得到了证明丝状噬菌体表面的乙肝表面抗原特异性多肽与单抗结合的酶联免疫吸附试验的支持。序列数据被用来绘制乙肝表面抗原的表面图，并推导出乙肝表面抗原101-207区的Cy-C痕迹的拓扑模型。对于晶体结构未知但可以获得一组具有代表性的单抗的其他蛋白质，这种方法应该是有用的。
The structure of the small hepatitis B virus surface antigen (HBsAg) was investigated by epitope mapping of four anti-HBsAg monoclonal antibodies (mAbs). Amino acid sequences of epitopes were derived from affinity-enrichment experiments (biopanning) using a filamentous phage peptide library. The library consists of 10(9) different clones bearing a 30-residue peptide fused to gene III. Sequence homologies between peptides obtained from panning the library against the antibodies and the native HBsAg sequence allowed for precise description of the binding regions. Three of four mAbs were found to bind to distinct discontinuous epitopes between amino acid residues 101 and 207 of HBsAg. The fourth mAb was demonstrated to bind to residues 121-124. The sequence data are supported by ELISA assays demonstrating the binding of the HBsAg-specific peptides on filamentous phage to mAbs. The sequence data were used to map the surface of HBsAg and to derive a topological model for the cy-carbon trace of the 101-207 region of HBsAg. The approach should be useful for other proteins for which the crystal structure is not available but a representative set of mAbs can be obtained.