| 妊娠特异性β1糖蛋白的分离纯化及特异抗血清的制备 |
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| 引用本文： | 吕懿娟 何赐科. 妊娠特异性β1糖蛋白的分离纯化及特异抗血清的制备[J]. 重庆医科大学学报, 1991, 16(2): 91-96 |
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| 作者姓名： | 吕懿娟 何赐科 |
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| 作者单位： | 重庆医科大学生化教研室(吕懿娟，何赐科，冯涛)，重庆医科大学生化教研室(赵瑜) |
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| 摘 要： | 本文采用硫酸铵盐析、DE-52离子交换层析、Sephadex G-200凝胶过滤及抗SP_1免疫亲和柱层析等方法从胎盘后血分离妊娠特异性β_1民糖蛋白(SP_1)所得SP_1纯化约207倍.纯化的SP_1与德国Behringwerke AG的SP_1抗血清在双向免疫扩散时有沉淀反应;免疫电泳时纯化SP_1移动的位置与β蛋白相当.聚丙烯酰胺凝胶电泳时纯化的SP_1呈现一条主带其分子量为90,000左右.免疫家免所得的抗血清经血浆蛋白吸收后得特异SP_1抗血清在双向免疫扩散时与正常男子或非孕妇女血清均无沉淀反应;与孕血反应形成的沉淀线与德国Behring werke AG的SP_1抗血清所形成的沉淀线相连;交义免疫电泳时呈现一个峰.此抗血清已成功地用于建立酶联免疫法测定SP_1含量
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| 关 键 词： | 妊娠特异性β_1糖蛋白 蛋白质纯化 |
PURIFI CATION OF PREGNANCY SPECIFIC B_1 GLYCOPROTEIN(SP_1)AND SPECIFIC ANTISERUM PREPARATION |
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| Lu Yijuan. PURIFI CATION OF PREGNANCY SPECIFIC B_1 GLYCOPROTEIN(SP_1)AND SPECIFIC ANTISERUM PREPARATION[J]. Journal of Chongqing Medical University, 1991, 16(2): 91-96 |
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| Authors: | Lu Yijuan |
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| Affiliation: | Lu Yijuan Department of Biochemistry |
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| Abstract: | The authers employed combinations of sequential (NH4)2 SO4 precipitation, DE-52 ion exchange chromatography,Sephadex G-200 gel filtration to isolate SP1 from retroplacental blood collected during labour. Anti-SP1 affinity chromatography has proven helpful in purification. The final product was shown about 207 fold of purification. Purified SP1 reacts with anti-SP1 serum from Behringwerke AG, migrates as p globulin in immunoelectrophoresis. SDS polyacrylamide gradient gel electroph-oresis indicates a major band with molecular weight around 90000. The antiserum obtained by immunization of rabbits with purified SP1 after appropriate absorption shows a single peak in cross immunoelectrophoresis, fails to react with serum from normal nonpregnant females and males, yields single line showing identity with SP1 antiserum from Behringwerke AG in immunodiffusion with pregnant plasma. This antiserum has been successfully used in establishing ELISA for SP1 |
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