小白鼠子宮分泌液自體抗原的純化輿特性鑑定

Abstract

利用小白雌鼠子宮分泌液(uterine fluid)免疫同系雌鼠，以酵素免疫分析法(ELISA)偵測，顯示可誘發雌鼠的自體免疫。由西方墨點法(Western blotting)知引起免疫反應的自體抗原其分子量為28kd，我們並將它命名為子宮液自體抗原(Uterine fluid autoantigen)，簡稱UFA。使用Sephadex G-100膠體過濾法和Reverse-phase-HPLC，我們將UFA分離。進一步的分析，發現UFA是一個近中性(PI約為7.6)的醣蛋白。其醣基約占醣蛋白重量的13%，而去醣後的UFA分子量降為23.5kd。我們並決定了UFA的氨基酸組成和N端的前十個氨基酸序到，而有關 UFA的完整一級結構及其所具的生理功能則尚需要進一步的研究才能知曉。

The female mice (8 wks) were autoimmunized with crude uterine fluid. The autoantisera were used for immunochemical detection of autoantigen(s) in uterine fluid. We found the autoantisera recognized a 28kd protein in uterine fluid and we named this protein uterine fluid autoantigen(UFA). We purified UFA through a series of procedures including Sephadex G-100 chromatography and Reverse-phase HPLC on a chemeco 5ODS-H column. UFA was shown to be a glycoprotein with isoelectric point of 7.6. The carbohydrate moiety constituted about 13% of total UFA mass. The protein core of UFA was estimated to be 23.5kd when glycoprotein was deglycosylated with TFMS. Moreover, we also determined the amino acid composition of UFA and the first ten amino acid sequences of its N-terminal region.