Lonsdale-Eccles J D, Grab D J
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
J Protozool. 1987 Nov;34(4):405-8. doi: 10.1111/j.1550-7408.1987.tb03201.x.
Bloodstream forms of African trypanosomes are routinely purified from blood components by a combination of centrifugation and chromatography on DEAE cellulose at pH 8.0. Here we report that the nonphysiological conditions used for DEAE chromatography of the parasites result in changes in the ATP levels of the trypanosomes and an enhanced release from the parasites of proteins such as variable surface glycoprotein, peptidase, and phospholipase. Some of these changes can be reduced by the addition of nucleosides to the elution buffer and, after the elution of the parasites, by immediate readjustment of the external pH to the normal physiological level of blood (pH 7.4).
非洲锥虫的血流型通常通过离心和在pH 8.0的DEAE纤维素上进行色谱分离相结合的方法从血液成分中纯化出来。我们在此报告,用于寄生虫DEAE色谱分析的非生理条件会导致锥虫ATP水平发生变化,并增强诸如可变表面糖蛋白、肽酶和磷脂酶等蛋白质从寄生虫中的释放。通过在洗脱缓冲液中添加核苷,以及在寄生虫洗脱后立即将外部pH值重新调整到血液的正常生理水平(pH 7.4),可以减少其中一些变化。