Laboratory of Experimental Optometry, Centre for Myopia Research, School of Optometry, Hong Kong Polytechnic University, Hong Kong, China.
State Key Laboratory of Chemical Biology and Drug Discovery, Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hong Kong, China.
Sci Data. 2021 Jan 26;8(1):27. doi: 10.1038/s41597-021-00813-1.
The retina is a key sensory tissue composed of multiple layers of cell populations that work coherently to process and decode visual information. Mass spectrometry-based proteomics approach has allowed high-throughput, untargeted protein identification, demonstrating the presence of these proteins in the retina and their involvement in biological signalling cascades. The comprehensive wild-type mouse retina proteome was prepared using a novel sample preparation approach, the suspension trapping (S-Trap) filter, and further fractionated with high-pH reversed phase chromatography involving a total of 28 injections. This data-dependent acquisition (DDA) approach using a Sciex TripleTOF 6600 mass spectrometer identified a total of 7,122 unique proteins (1% FDR), and generated a spectral library of 5,950 proteins in the normal C57BL/6 mouse retina. Data-independent acquisition (DIA) approach relies on a large and high-quality spectral library to analyse chromatograms, this spectral library would enable access to SWATH-MS acquisition to provide unbiased, multiplexed, and quantification of proteins in the mouse retina, acting as the most extensive reference library to investigate retinal diseases using the C57BL/6 mouse model.
视网膜是一种关键的感觉组织，由多个细胞层组成，这些细胞层协同工作，以处理和解码视觉信息。基于质谱的蛋白质组学方法允许高通量、非靶向的蛋白质鉴定，证明了这些蛋白质在视网膜中的存在及其在生物信号级联中的参与。使用一种新颖的样品制备方法，悬浮捕获（S-Trap）过滤器，进一步用高 pH 反相色谱法对全野生型小鼠视网膜蛋白质组进行了分离，总共进行了 28 次注射。使用 Sciex TripleTOF 6600 质谱仪的这种数据依赖型采集（DDA）方法总共鉴定了 7122 种独特的蛋白质（1% FDR），并在正常 C57BL/6 小鼠视网膜中生成了 5950 种蛋白质的光谱库。数据非依赖型采集（DIA）方法依赖于大型和高质量的光谱库来分析色谱图，该光谱库将能够进行 SWATH-MS 采集，以提供对小鼠视网膜中蛋白质的无偏、多重和定量分析，作为使用 C57BL/6 小鼠模型研究视网膜疾病的最广泛参考库。
