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制备用于蛋白质组学研究分析的冷沉淀和冷沉淀去除血浆。

Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Research Analysis.

机构信息

Transfusion Science, Melbourne, VIC, Australia.

School of Public Health and Preventive Medicine, Monash University, Melbourne, VIC, Australia.

出版信息

Methods Mol Biol. 2023;2628:41-49. doi: 10.1007/978-1-0716-2978-9_4.

Abstract

When frozen plasma is slowly thawed in cold conditions (1-6 °C), high-molecular-weight plasma proteins precipitate forming a concentrate known as cryoprecipitate. The concentrate is enriched with several important coagulation proteins, including fibrinogen, antihemophilic factor (factor VIII), von Willebrand factor, fibrin stabilizing factor (factor XIII), fibronectin, and small amounts of other plasma proteins. In current medical practice, clinical-grade preparations of cryoprecipitate are used mostly to correct fibrinogen deficiency caused by acute blood loss or due to functional abnormalities of the fibrinogen protein. In the past, cryoprecipitate was used to treat von Willebrand disease and hemophilia A (factor VIII deficiency), but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies. Cryo-depleted plasma (also called cryosupernatant) is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma and contains all the remaining soluble plasma proteins. This protocol describes the research-scale preparation of cryoprecipitate and cryo-depleted plasma suitable for proteomic studies and is based on the procedures used to prepare clinical-grade cryoprecipitate.

摘要

当冷冻血浆在低温条件下(1-6°C)缓慢解冻时,高分子量血浆蛋白沉淀形成一种浓缩物,称为冷沉淀。该浓缩物富含几种重要的凝血蛋白,包括纤维蛋白原、抗血友病因子(因子 VIII)、血管性血友病因子、纤维蛋白稳定因子(因子 XIII)、纤维连接蛋白和少量其他血浆蛋白。在当前的医学实践中,临床级别的冷沉淀制剂主要用于纠正因急性失血或纤维蛋白原蛋白功能异常引起的纤维蛋白原缺乏症。过去,冷沉淀曾用于治疗血管性血友病和甲型血友病(因子 VIII 缺乏症),但更高度纯化的凝血因子浓缩物或重组蛋白制剂的可用性已经取代了冷沉淀在这些凝血疾病中的应用。去冷沉淀血浆(也称为冷沉淀上清液)是从冷冻解冻的血浆中去除冷沉淀后剩余的血浆上清液,含有所有剩余的可溶性血浆蛋白。本方案描述了适合蛋白质组学研究的研究规模的冷沉淀和去冷沉淀血浆的制备,该方案基于制备临床级冷沉淀的程序。

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