蛋白质氯胺-T双相碘标法的建立及其应用

常规的蛋白质碘标方法易引起被标细胞因子的失活，是受体配基竞争结合实验失败的原因之一.试用氯胺-T双相碘标法标记rhG-CSF和rhEPO，并应用受体配基竞争结合分析法测定NFS-60细胞G-CSF受体及BET-2细胞EPO受体的特性.结果显示所获 ¹²⁵I-EPO和 ¹²⁵I-G-CSF放射比活度均较高；发现BET-2细胞有高、低两种亲和力的EPO受体，NFS-60细胞只有一种高亲和力的G-CSF受体，所获结果与文献资料相一致.说明氯胺-T双相碘标法是细胞因子同位素碘标记的理想方法之一.

Establishment of Methods for Two-phase Iodination of Chromine-T and Applying in Cytokine Receptor Studies

The routine iodination methods are easy to lead to biological inactivation of cytokines, which is one of reasons for failure of ligand receptor binding assay. By using two-phase iodination of chromine-T, rhG-CSF and rhEPO were iodinated with Na¹²⁵I and the characteristic of EPO receptor in BET-2 cell and G-CSF receptor in NFS-60 cell were observed by receptor ligand binding assay. It was showed that the radioactivity of ¹²⁵I-G-CSF and ¹²⁵I-EPO were much higher than that obtained by other methods and it was found that there are high and low affinity EPO receptors in BET-2 cells and one high affinity G-CSF receptor in NFS-60 cells. These results are in accord with literature data. It suggests that the two-phase iodination of chromine-T is an ideal method to iodinate cytokines.