内生致热原的纯化实验研究

Studies on the purification of rabbit monocyte pyrogen

本实验采用家兔单核细胞致热原(monocytic endogenous pyrogen)粗制品,通过超滤、Sephadex G75、QAE-Sephadex A50和等电聚焦电泳方法进一步纯化内生致热原。实验结果表明:凝胶层析纯化内生致热原,有两个致热原性蛋白吸收峰,其致热活性0.3μg蛋白和5.0μg蛋白均引起0.6℃的发热效应。等电聚焦电泳获得的致热原活性蛋白质的等电点,分别为5.1和3.6。

Crude monocytic endogenous pyrogens from rabbits' blood were further purified by means of ultrafiltration membranes. The sterile pyrogen obtained was passed through gel filtration on a 1.5 by 100cm G-75 Sephadex column, and subje- cted to a 1.5 by 60cm ion-exchange chromatography on QAE Sephadex or subjected to isoelectric focusing. Under our experimental conditions high concentration of monocyte pyrogen could be obtained. In above described method used two pyrogen peak whose specific activity was 0.5—3.0ug, caused fever up to 0.6℃ in rabbits. In addition, isoelectric focusing column employed indicated isoeletric points of specific activity protein that induced fever were PI 5.1 and 3.6 respectively.