【摘要】 蛋白质力谱测试是研究生物学的重要组成部分.蛋白质力谱测试的成功取决于两大技术:一是分子尺度的力谱测量能力,即皮牛量级的力学分辨力和纳米级别的空间分辨力;二是蛋白质样品制备能力,通过将微观生物分子链与介观微球/探针/基底耦联,实现对分子尺度蛋白质的有效测量.然而,蛋白质样品的耦联情况很难在视觉上直接观测,耦联效果的好坏决定测试的成败.因此,蛋白质样品相关制备方法、耦联工艺一直是单分子力谱测试中的研究重点.针对3种主要单分子力谱测试技术即光镊、磁镊及原子力显微术对测试蛋白质的需求特点,特别是对多分子链耦合样品的测试需求特点,介绍了多种基于基底修饰、蛋白质修饰和DNA链修饰的提高连接待测蛋白质与DNA链/微球/探针/玻片/云母片的方法,分析了各自的优缺点,并总结了典型的应用,为相关领域中样品的制备提供参考方案.更多还原

【Abstract】 Protein force spectroscopy testing is one of the important components of the study of biology. The success of protein force spectroscopy testing depends on the following factors:the capability of force spectroscopy measurement at the molecular scale,namely piconewton-scale force resolution and nanometer-scale spatial resolution,and the protein sample preparation technique,which achieves effective measurement of protein samples at the molecular scale through the coupling of microscopic biomolecular chains with mesoscopic microspheres/probes/substrates. However,the coupling of protein samples is invisible and its coupling effect seriously affects the test efficiency.Therefore,the preparation method and coupling technology of protein samples are the key points in the measurement of single-molecule force spectroscopy. According to the various demands of three single-molecule force spectroscopy testing techniques on the samples,particularly the test requirement of multi-molecular chain-coupled samples,the authors introduce different methods to improve the connection efficiency between protein and DNA handles/microspheres/probes/glass slides/mica in optical tweezers,magnetic tweezers,and atomic force microscopy based on substrate,protein,and DNA modification,and discuss their advantages and disadvantages. The typical applications are summarized to provide a reference for the preparation of biological samples in related fields.更多还原