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尿激酶抗人交联纤维蛋白-D-二聚体单抗化学偶合体的制备

Preparation of Conjugate of Urokinase With Monoclonal Antibody against Fragment D dimer of Human Cross linked Fibrin

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【作者】 欧阳伟民余瑞元徐长法

【Author】 Quyang Wei Min Yu Rui Yuan Xu Chang Fa (Department of Biochemistry and Molecular Biology,Peking University,Beijing 100871)

【机构】 北京大学生物化学及分子生物学系

【摘要】 利用双功能试剂N-琥珀酰亚胺-3(2-二硫吡啶)丙酸酯(SPDP)作交联剂,合成了尿激酶(UK)-抗人交联纤维蛋白降解物D-二聚体单抗(MA-HID1)化学偶合体(UKMA-HID1),并用苯甲脒-Sepharose6B及人交联纤维蛋白降解物D-二聚体-Sepharose4B亲和柱纯化,获得偶合体产物.SDS-PAGE呈现一条带,其分子量约为200000.纤维蛋白平板法测活结果显示,偶合体中酶比活为53000IU/mg尿激酶蛋白,与偶联前的54300IU/mg蛋白相仿.ELISA测试显示,偶合体对人交联纤维蛋白降解物D-二聚体有免疫反应性,并且与偶联前的抗D-二聚体单抗对此抗原的反应性相当

【Abstract】 Urokinase (UK) and a murine monoclonal antibody(MA HID 1) directed against fragment D dimer of human cross linked fibrin were conjugated,using the cross linked agent N Succinimidyl 3(2 pyridylidithio)propionate(SPDP).The conjugate(UK·MA HID 1)was purified by affinity chromatography on Benzamidine Sepharose 6B and fragment D dimer Sepharose 4B columns.On SDS PAGE,it migrated as a main band with apparent M r of 200 000 .Specific activity on fibrin plates was 53 000 IU/mg urokinase protein as compared to 54 000 IU/mg protein for urokinase protein before conjugation.The results of the ELISA showed that the conjugate had immuno reactivity for fragment D dimer,and similar to reactivity of MA HID 1 for this antigen.

【基金】 国家863计划
  • 【分类号】Q78
  • 【被引频次】3
  • 【下载频次】83
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