Institute of Biomedical Chemistry, 119121 Moscow, Russia.
Int J Mol Sci. 2021 Jan 4;22(1):431. doi: 10.3390/ijms22010431.
An approach to highly-sensitive mass spectrometry detection of proteins after surface-enhanced concentrating has been elaborated. The approach is based on a combination of mass spectrometry and atomic force microscopy to detect target proteins. (1) Background: For this purpose, a technique for preliminary preparation of molecular relief surfaces formed as a result of a chemical or biospecific concentration of proteins from solution was developed and tested on several types of chip surfaces. (2) Methods: mass spectrometric identification of proteins using trailing detectors: ion trap, time of flight, orbital trap, and triple quadrupole. We used the electrospray type of ionization and matrix-assisted laser desorption/ionization. (3) Results: It is shown that when using locally functionalized atomically smooth surfaces, the sensitivity of the mass spectrometric method increases by two orders of magnitude as compared with measurements in solution. Conclusions: It has been demonstrated that the effective concentration of target proteins on specially prepared surfaces increases the concentration sensitivity of mass spectrometric detectors-time-of-flight, ion trap, triple quadrupole, and orbital ion trap in the concentration range from up to 10 M.
已经阐述了一种用于表面增强浓缩后蛋白质的高灵敏度质谱检测的方法。该方法基于质谱和原子力显微镜的结合,用于检测靶蛋白。(1)背景:为此,开发了一种用于初步制备分子浮雕表面的技术,该表面是通过从溶液中化学或生物特异性浓缩蛋白质而形成的,并在几种类型的芯片表面上进行了测试。(2)方法:使用尾随探测器进行蛋白质的质谱鉴定:离子阱、飞行时间、轨道阱和三重四极杆。我们使用电喷雾类型的电离和基质辅助激光解吸/电离。(3)结果:结果表明,当使用局部功能化的原子平滑表面时,与在溶液中的测量相比,质谱方法的灵敏度提高了两个数量级。结论:已经证明,在专门制备的表面上靶蛋白的有效浓度增加了质量光谱检测器的浓度灵敏度-飞行时间,离子阱,三重四极杆和轨道离子阱在 10M 范围内的浓度。