水性和基于分子质量的分离方法,用于从哺乳动物总细胞蛋白中发现自身抗体
Mirei Date1, Ai Miyamoto1, Tomoko Honjo1
1Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, Japan.
Protein science : a publication of the Protein Society
|August 28, 2023
概括
这项研究引入了一种可复制,可扩展的蛋白质组学方法,用于发现自身抗体生物标志物. 新的二维凝电泳系统有效地分析细胞总蛋白质,用于对自身免疫和瘤免疫的免疫监测.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 免疫学 免疫学 免疫学
- 生物标志物发现发现
背景情况:
- 血清自身抗体简介对于理解自身免疫和瘤免疫至关重要.
- 识别自身抗体生物标志物对于开发针对免疫相关疾病的免疫监测系统至关重要.
研究的目的:
- 为大规模的自身抗体发现提供一种实用,可复制和可扩展的方法.
- 建立一种新的二维凝电泳系统,用于基于蛋白质学的自身抗体识别.
主要方法:
- 利用来自哺乳动物细胞的无核酸,变质,水溶性总细胞蛋白.
- 采用逆相高性能液态染色学 (HPLC) 基于疏水性进行蛋白质分离.
- 应用二维凝电泳,包括二甲基硫酸聚烯胺凝电泳 (SDS-PAGE),用于分子质量分析,免疫涂抹和液体染色学-质谱/质谱 (LC-MS/MS).
主要成果:
- 证明了准备的同质,变质的总细胞蛋白质在精确分离方面的优越性.
- 证实了所有程序的高可重现性和可扩展性,从蛋白质提取到SDS-PAGE.
- 成功地利用二维凝电泳系统进行免疫涂抹和LC-MS/MS分析.
结论:
- 新的二维凝电泳系统是一个可行的基于蛋白质组学的替代方法,用于大规模的自身抗体发现.
- 该方法提供了一个强大的平台,用于识别对免疫监测至关重要的自身抗体生物标志物.
更多相关视频
11:09An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
Published on: April 17, 2018
10.2K
09:32Immunopeptidomics: Isolation of Mouse and Human MHC Class I- and II-Associated Peptides for Mass Spectrometry Analysis
Published on: October 15, 2021
12.3K
相关概念视频
01:20Overview Of Cell Separation And Isolation
Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
01:20Immunoprecipitation
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
01:15Western Blotting
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.