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基质辅助激光解吸电离质谱法检测体内吸附在隐形眼镜上的蛋白质。

Matrix-assisted laser desorption ionization mass spectrometry detection of proteins adsorbed in vivo onto contact lenses.

作者信息

Kingshott P, St John H A, Chatelier R C, Griesser H J

机构信息

CSIRO Molecular Science, Private Bag 10, Clayton South MDC, Clayton 3169, Australia.

出版信息

J Biomed Mater Res. 2000 Jan;49(1):36-42. doi: 10.1002/(sici)1097-4636(200001)49:1<36::aid-jbm5>3.0.co;2-b.

Abstract

Identification of the biomolecules that form the first adsorbed monolayer, which thus effect "interface conversion", in competitive adsorption from multicomponent biological solutions can be challenging because of limitations in mass resolution and sensitivity of established techniques. In this study matrix-assisted laser desorption ionization (MALDI) time of flight mass spectrometry is developed and applied as a novel surface analytical method to enable analysis of adsorbed multicomponent biomolecule layers directly on the biomaterial surfaces. We show that proteins adsorbed in vivo (on human eyes) on contact lenses can be detected rapidly and conveniently by the diagnostic highly resolved mass signals recorded by MALDI mass spectrometry. This new approach allows detection of minor (and major) proteinaceous constituents of biofouled layers at levels substantially below monolayer coverage. Identification is done by comparison with molecular masses of known proteins. Specifically, it is shown that in addition to lysozyme, other low molecular weight proteins adsorb from human tear fluid onto contact lenses; these proteins had not been detected in earlier studies using other techniques.

摘要

从多组分生物溶液中进行竞争性吸附时,识别形成首个吸附单分子层从而实现“界面转化”的生物分子可能具有挑战性,因为现有技术在质量分辨率和灵敏度方面存在局限性。在本研究中,基质辅助激光解吸电离(MALDI)飞行时间质谱被开发并用作一种新型表面分析方法,以能够直接在生物材料表面分析吸附的多组分生物分子层。我们表明,通过MALDI质谱记录的诊断性高分辨率质量信号,可以快速方便地检测出体内(在人眼上)吸附在隐形眼镜上的蛋白质。这种新方法能够在远低于单分子层覆盖水平的情况下检测生物污染层中的微量(和主要)蛋白质成分。通过与已知蛋白质的分子量进行比较来进行鉴定。具体而言,研究表明,除溶菌酶外,其他低分子量蛋白质也会从人泪液吸附到隐形眼镜上;在早期使用其他技术的研究中未检测到这些蛋白质。

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