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蛋白质组学研究用冷沉淀和冷沉淀去除血浆的制备方案。

A Protocol for the Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Studies.

作者信息

Sparrow Rosemary L, Simpson Richard J, Greening David W

机构信息

Transfusion Science, Melbourne, VIC, Australia.

Department of Immunology and Pathology, Monash University, AMREP, 89 Commercial Road, Melbourne, VIC, 3004, Australia.

出版信息

Methods Mol Biol. 2017;1619:23-30. doi: 10.1007/978-1-4939-7057-5_2.

Abstract

Cryoprecipitate is a concentrate of high-molecular-weight plasma proteins that precipitate when frozen plasma is slowly thawed at 1-6 °C. The concentrate contains factor VIII (antihemophilic factor), von Willebrand factor (vWF), fibrinogen, factor XIII, fibronectin, and small amounts of other plasma proteins. Clinical grade preparations of cryoprecipitate are mainly used to treat fibrinogen deficiency caused by acute bleeding or 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but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies.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 ("cryosupernatant") is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma. It contains all the other plasma proteins and clotting factors present in plasma that remain soluble during cold-temperature thawing of the plasma. This protocol describes the clinical-scale preparation of cryoprecipitate and cryo-depleted plasma for proteomic studies.

摘要

冷沉淀是一种高分子量血浆蛋白浓缩物,当冷冻血浆在1-6°C缓慢解冻时会沉淀。该浓缩物含有因子VIII(抗血友病因子)、血管性血友病因子(vWF)、纤维蛋白原、因子XIII、纤连蛋白和少量其他血浆蛋白。临床级冷沉淀制剂主要用于治疗急性出血引起的纤维蛋白原缺乏或纤维蛋白原蛋白功能异常。过去,冷沉淀用于治疗血管性血友病和甲型血友病(因子VIII缺乏),但现在有了更高纯度的凝血因子浓缩物或重组蛋白制剂,冷沉淀已不再用于这些凝血障碍的治疗。冷沉淀去除后的血浆上清液(“冷上清液”)是从冻融血浆中去除冷沉淀后剩余的血浆上清液。它包含血浆中所有其他血浆蛋白和凝血因子,这些物质在血浆低温解冻过程中仍保持可溶状态。本方案描述了用于蛋白质组学研究的临床规模冷沉淀和冷沉淀去除后血浆的制备方法。

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