Evaluation of the molecular mechanisms by which an antibody recognizes a specific antigen could help in better understanding of the protein recognition mechanisms. We previously showed that anti-hen egg lysozyme (HEL) monoclonal antibody, HyC1, recognized the structural and hydrodynamic change in HEL. Here, we generated HyC1 single-chain Fv (scFv), and characterized it using different structural and biophysical methods. Similar to HyC1 monoclonal antibody, HyC1 scFv could recognize native HEL from carboxymethylated Cys6 and Cys127 HEL (CM6,127-HEL). Comparison of the binding thermodynamics of HyC1 scFv between HEL and CM6,127-HEL showed that the binding enthalpy change was different, while the binding entropy was remained unchanged. The results indicated that the fluctuation of the residual native structure in both HEL and CM6,127-HEL was similar. The NMR experiments for 15N-labeled HyC1 scFv indicated that the flexibility of HyC1 scFv decreased upon the binding to HEL.

中文翻译：

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单链Fv抗体HyC1的结构和功能评估，可识别鸡蛋溶菌酶的残留天然结构。

抗体识别特定抗原的分子机制的评估可以帮助更好地理解蛋白质识别机制。我们以前显示抗鸡卵溶菌酶（HEL）单克隆抗体HyC1识别HEL中的结构和流体动力学变化。在这里，我们生成了HyC1单链Fv（scFv），并使用不同的结构和生物物理方法对其进行了表征。与HyC1单克隆抗体相似，HyC1 scFv可以从羧甲基化的Cys6和Cys127 HEL（CM6,127-HEL）识别天然HEL。HEL和CM6,127-HEL之间HyC1 scFv的结合热力学比较表明，结合焓变化不同，而结合熵保持不变。结果表明，HEL和CM6,127-HEL中残留天然结构的波动相似。