Muscovite mica with an amino silane-modified surface is commonly used as a substrate in atomic force microscopy (AFM) studies of biological macromolecules. Herein, the efficiency of two different protein immobilization strategies employing either (N-hydroxysuccinimide ester)-based crosslinker (DSP) or benzophenone-based photoactivatable crosslinker (SuccBB) has been compared using AFM and mass spectrometry analysis. Two proteins with different physicochemical properties—human serum albumin (HSA) and horseradish peroxidase enzyme protein (HRP)—have been used as model objects in the study. In the case of HRP, both crosslinkers exhibited high immobilization efficiency—as opposed to the case with HSA, when sufficient capturing efficiency has only been observed with SuccBB photocrosslinker. The results obtained herein can find their application in commonly employed bioanalytical systems and in the development of novel highly sensitive chip-based diagnostic platforms employing immobilized proteins. The obtained data can also be of interest for other research areas in medicine and biotechnology employing immobilized biomolecules.

中文翻译：

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用光交联剂将共价蛋白固定在白云母云母表面

具有氨基硅烷修饰表面的白云母通常用作生物大分子的原子力显微镜（AFM）研究的底物。在本文中，使用原子力显微镜（AFM）和质谱分析法比较了使用（N-羟基琥珀酰亚胺酯）基交联剂（DSP）或二苯甲酮基光活化交联剂（SuccBB）的两种不同蛋白质固定策略的效率。两种具有不同理化特性的蛋白质-人血清白蛋白（HSA）和辣根过氧化物酶蛋白（HRP）-已被用作研究的模型对象。在HRP的情况下，两种交联剂均显示出高固定化效率，而HSA则相反，而只有在SuccBB光交联剂中才能观察到足够的捕获效率。本文获得的结果可在常用的生物分析系统中以及在使用固定化蛋白质的新型高灵敏度基于芯片的诊断平台的开发中找到其应用。所获得的数据也可能对采用固定化生物分子的医学和生物技术的其他研究领域感兴趣。