摘要
目的:探讨血管内皮细胞生长因子(VEGF)放射性碘标记的方法。方法:采用肝素亲和层析的方法,纯化^(125)I-VEGF。通过SDS-PAGE进行同质性分析。利用VEGF能够与内皮细胞特异性结合的特点,鉴定其生物学活性。结果:^(125)I-VEGF的比活性为(1.6～1.9)×10~5cpm/ng。12%SDS-PAGE表明在未还原和还原状态下^(125)IVEGF在42ku和21ku处显示单一带。^(125)I-VEGF与内皮细胞能够特异地结合,并能够被TSP抑制。结论:肝素亲和层析的方法能够有效地纯化^(125)I-VEGF,并且具有生物学活性。
Objective:To explore the method of iodination of vascular endothelial growth factor (VEGF) . Methods:125 I-VEGF was purified with heparin affinity chromatography. Homogeneity of 125 I-VEGF was described by SDS-PAGE. Analysis of its bio-logical activity was investigated by specific binding of 125 I-VEGF and ECV-304 cells. Results:The specific activity of 125 I-VEGF ranged from (1.6~1.9)×105 cpm/ng. 12% SDS-PAGE showed that the iodinated VEGF migrated as a single band of 42 ku,reduction with β-mercapto ethanol caused the dissociation of the dimer into 21 ku polypeptides. Binding of 125 I-VEGF and ECV-304 cells was specific and inhibited by thrombospondin (TSP) . Conclusion:Heparin affinity chromato-graphy could efficiently purify 125 I-VEGF which was biologically active.
出处
《天津医药》
CAS
2000年第3期153-155,共3页
Tianjin Medical Journal
基金
中国医学科学院基金
