Eye Pathology Section, Department of Neuroscience and Pharmacology, University of Copenhagen, Copenhagen, Denmark.
Department of Medical Biochemistry, Aarhus University, Aarhus, Denmark.
Biol Proced Online. 2009 Dec 24;12(1):56-88. doi: 10.1007/s12575-009-9019-7.
The intention of this review is to provide an overview of current methodologies employed in the rapidly developing field of ocular proteomics with emphasis on sample preparation, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Appropriate sample preparation for the diverse range of cells and tissues of the eye is essential to ensure reliable results. Current methods of protein staining for 2D-PAGE, protein labelling for two-dimensional difference gel electrophoresis, gel-based expression analysis and protein identification by MS are summarised. The uses of gel-free MS-based strategies (MuDPIT, iTRAQ, ICAT and SILAC) are also discussed. Proteomic technologies promise to shed new light onto ocular disease processes that could lead to the discovery of strong novel biomarkers and therapeutic targets useful in many ophthalmic conditions.
本综述的目的是概述眼部蛋白质组学这一快速发展领域中当前使用的方法学，重点是样品制备、二维聚丙烯酰胺凝胶电泳（2D-PAGE）和质谱（MS）。适当的样品制备对于眼睛的各种细胞和组织至关重要，以确保可靠的结果。本文总结了当前用于 2D-PAGE 的蛋白质染色方法、二维差异凝胶电泳的蛋白质标记、基于凝胶的表达分析以及 MS 进行蛋白质鉴定的方法。还讨论了无胶 MS 策略（MuDPIT、iTRAQ、ICAT 和 SILAC）的应用。蛋白质组学技术有望为眼部疾病过程提供新的见解，从而发现对许多眼科疾病有用的强新型生物标志物和治疗靶点。
