纯化血红蛋白的简便方法。

Convenient method to purify hemoglobin.

作者信息

Sakai H, Takeoka S, Nishide H, Tsuchida E

机构信息

Department of Polymer Chemistry, Waseda University, Tokyo, Japan.

出版信息

Artif Cells Blood Substit Immobil Biotechnol. 1994;22(3):651-6. doi: 10.3109/10731199409117895.

DOI:10.3109/10731199409117895

PMID:7994386

Abstract

A convenient method to purify Hb solution from outdated RBC has been established for the starting material of Hb-based blood substitutes. To prevent MetHb formation during the procedure, Hb in RBC was carbonylated in advance. Then RBC was mixed with organic solvent for hemolysis and centrifuged for removal of stroma. The resulting SFHb solution was heated at 60 degrees C and generated precipitates were removed out by centrifugation. The purity of Hb (25 g/dl) was confirmed by SDS-PAGE. IEF and oxygen binding property of the Hb solution also guaranteed its purity and no denaturation of Hb. This method is applicable to large scale production of the purified Hb for the starting material of Hb-based blood substitutes.

摘要

已建立一种从过期红细胞中纯化血红蛋白（Hb）溶液的简便方法，作为基于血红蛋白的血液替代品的起始原料。为防止该过程中高铁血红蛋白（MetHb）的形成，红细胞中的Hb预先进行了羰基化处理。然后将红细胞与有机溶剂混合进行溶血，并通过离心去除基质。所得的无基质血红蛋白（SFHb）溶液在60℃加热，通过离心去除产生的沉淀。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）确认了Hb（25 g/dl）的纯度。Hb溶液的等电聚焦（IEF）和氧结合特性也保证了其纯度以及Hb未发生变性。该方法适用于大规模生产纯化的Hb，作为基于血红蛋白的血液替代品的起始原料。