Lai Andrew, Kinhal Vyjayanthi, Nuzhat Zarin, Menon Ramkumar, Rice Gregory E, Salomon Carlos
Exosome Biology Laboratory, Centre for Clinical Diagnostics, University of Queensland Centre for Clinical Research, Royal Brisbane and Women's Hospital, The University of Queensland, Bldg. 71/918, Royal Brisbane Hospital, Brisbane, QLD, 4029, Australia.
Division of Maternal-Fetal Medicine & Perinatal Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA.
Methods Mol Biol. 2018;1710:139-153. doi: 10.1007/978-1-4939-7498-6_11.
Exosomes are membrane-bound nanovesicles that transport molecular signals (e.g., proteins) between cells and are released from a wide range of cells, including the human placenta. Interestingly, the levels of exosomes present in maternal circulation are higher in preeclamptic pregnancies and their protein content profile change in response to the microenvironment milieu. Through the discovery of candidate biomarkers, mass spectrometry (MS)-based proteomics may provide a better understanding of the pathophysiology underlying pregnancy-associated disorders. With advances in sample preparation techniques, computational methodologies, and bioinformatics, MS-based proteomics have addressed the challenge of identifying and quantifying thousands of proteins and peptides from a variety of complex biological samples. Despite increasing interest in biomarker diagnostics, the complex nature of biological matrices (e.g., plasma) poses a challenge for candidate biomarker discovery. Here we describe a workflow to prepare exosomes for proteomic analysis.
外泌体是膜结合的纳米囊泡，可在细胞间传递分子信号（如蛋白质），并由包括人胎盘在内的多种细胞释放。有趣的是，子痫前期妊娠母体循环中外泌体的水平较高，并且其蛋白质含量谱会根据微环境而变化。通过发现候选生物标志物，基于质谱（MS）的蛋白质组学可能有助于更好地理解妊娠相关疾病的病理生理学。随着样品制备技术、计算方法和生物信息学的进步，基于MS的蛋白质组学已经解决了从各种复杂生物样品中鉴定和定量数千种蛋白质和肽的挑战。尽管人们对生物标志物诊断的兴趣日益增加，但生物基质（如血浆）的复杂性质给候选生物标志物的发现带来了挑战。在此，我们描述了一种用于蛋白质组学分析的外泌体制备工作流程。
