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首页> 外文期刊>The FEBS journal >Regional and segmental flexibility of antibodies in interaction with antigens of different size
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Regional and segmental flexibility of antibodies in interaction with antigens of different size

机译：抗体与不同大小抗原相互作用的区域和分段灵活性

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摘要

The interaction of antibodies (Abs) with protein antigens (Ags) of different size, such as hen egg white lysozyme, ovalbumin, and bovine serum albumin, was examined using analytical ultracentrifugation, electrospray ionization time-of-flight mass spectrometry, and surface plasmon resonance in order to estimate regional and segmental Ab flexibility. When both Abs and Ags were free in solution, sedimentation equilibrium and surface plasmon resonance analyses showed the formation of an Ag(2)Ab(1) complexes regardless of Ag size, suggesting that the Fab arms were able to move to avoid interference between Ags bound to Ab combining sites. The Ag(2)Ab(1) complex, as well as the Ag(2)Ab(1) complex, was observed by MS. However, when Abs were immobilized on the surface of a sensor chip through the Fc region, the stoichiometry of the Ag-Ab complex was dependent on the Ag size; Ag(2)Ab(1) forming with hen egg white lysozyme and Ag(2)Ab(1) with ovalbumin and bovine serum albumin. These results indicated that immobilization of the Fc region reduces the dynamic range of the Fab arms and results in interference from the first Ag bound to either combining site, which in turn prevents the binding of the second Ag to the other combining site. Our results allow Lis to propose that the Fab arms of B-cell receptors whose Fc regions are immobilized on cell surface have a reduced dynamic range.
机译：使用分析超速离心，电喷雾电离飞行时间质谱和表面等离激元分析了抗体（Abs）与不同大小的蛋白质抗原（Ags）的相互作用，例如蛋清溶菌酶，卵清蛋白和牛血清白蛋白。共振以估计区域和分段Ab的灵活性。当Abs和Ags都游离在溶液中时，沉淀平衡和表面等离子体共振分析表明无论Ag大小如何，均会形成Ag（2）Ab（1）复合物，这表明Fab臂能够移动以避免Ags之间的干扰绑定到Ab组合站点。质谱观察到的Ag（2）Ab（1）配合物，以及Ag（2）Ab（1）配合物。但是，当通过抗体区域将Abs固定在传感器芯片表面时，Ag-Ab配合物的化学计量取决于Ag的大小。 Ag（2）Ab（1）与鸡蛋清溶菌酶形成，Ag（2）Ab（1）与卵清蛋白和牛血清白蛋白形成。这些结果表明，Fc区的固定减少了Fab臂的动态范围，并导致来自结合至任一结合位点的第一Ag的干扰，这继而阻止了第二Ag结合至另一结合位点。我们的结果允许Lis提出其Fc区固定在细胞表面上的B细胞受体的Fab臂具有减小的动态范围。

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