Construction and Characterization of Novel Staphylokinase Variants with Antiplatelet Aggregation Activity and Reduced Immunogenecity
Construction and Characterization of Novel Staphylokinase Variants with Antiplatelet Aggregation Activity and Reduced Immunogenecity作者机构：Department of Molecular Genetics Shanghai Medical School Fudan University Key Laboratory of Molecular Medicine Ministry of Education Shanghai 200032 China
出 版 物：《Acta Biochimica et Biophysica Sinica》 (生物化学与生物物理学报(英文版))
年 卷 期：2004年第36卷第5期
页 面：336-342页
核心收录：
学科分类：1001[医学-基础医学(可授医学、理学学位)] 10[医学]
主 题：staphylokinase RGD antiplatelet aggregation immunogenicity
摘 要：To develop target thrombolytic agents with fibrinolytic activity, antiplatelet aggregation activity and reduced immunogenicity, two staphylokinase variants containing Arg-Gly-Asp (RGD) motif were constructed. Gene expression was induced in E. coli JF1125 and the variants, designated DGR and RL1, were purified with gel filtration and ion-exchange chromatography and the purity was over 95%. The fibrinolytic activity and kinetic constants of the two variants were comparable to those of recombinant wild-type staphylokinase. Both the variants can inhibit the platelet aggregation at a final concentration of 2 μM. The titers of antibodies against variants were much lower than those against recombinant staphylokinase in guinea pigs, which indicated that the immunogenicity of the variants was greatly reduced. These results confirm that it is possible to design and produce a bifunctional protein that possesses fibrinolytic and antiplatelet aggrega- tion activities.
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