mPEG修饰红细胞Rh抗原稳定性探究.docVIP

mPEG修饰红细胞Rh抗原稳定性探究 　 作者：邱艳，查，檀英霞，章扬培 【摘要】 本研究旨在探究mPEG修饰的红细胞Rh抗原的稳定性。利用红细胞膜蛋白SDS电泳技术分析mPEG修饰后红细胞膜蛋白与mPEG结合情况，用红细胞血影凝集实验及4℃ CPD保养液保存的修饰红细胞与匹配血液体外混血实验，观察mPEG修饰后红细胞Rh抗原被遮蔽的稳定性。结果发现：不同保存时间的mPEG修饰的红细胞在模拟输血（即混血前后）的血型保持不变，同时mPEG修饰的红细胞在保存过程中游离血红蛋白水平正常，并且混血后经37℃孵育的mPEG修饰的红细胞游离血红蛋白水平低于不混合的修饰的红细胞。比较分析PEG特异的碘染和考马斯亮蓝染的显色图谱发现，特殊的碘染显示出mPEG与红细胞膜蛋白结合的条带，mPEG-SPA与红细胞膜蛋白结合后导致蛋白分子迁移速度发生改变。mPEG修饰的红细胞血影凝集实验证实，修饰红细胞在质膜裂损后 mPEG仍有效地遮蔽抗原。结论：mPEG-SPA不论在红细胞膜蛋白被单独提取后，还是膜破损后以及存活状态下，均能与红细胞膜蛋白稳固结合。 【关键词】 mPEG修饰红细胞；Rh抗原；红细胞 　　Rh Antigen Stability of mPEG Modified Red Blood Cells 　　AbstractThe objective of study was to investigate the Rh antigen stability of mPEG-modified RBC. RBC membrane protein SDStechnology was used to analyze the combination of the mPEG modified RBC membrane protein with mPEG molecules; the RBC ghost coagulation test and 4℃ CPD-preserved modified RBC mixed with matched blood were used to observe the stability of RBC Rh antigen camouflaged by mPEG. The results showed that the blood groups of stored mPEG-modified RBC were kept consistency before or after simulating transfusion, i.e. mixture of modified RBC with matched bloods, while the plasma hemoglobin after simulating transfusion was not only within the normal range during the storage, but also less than that before simulating transfusion even after incubation at 37℃. The electrophoresis pattern stained with iodine and Coomassie blue displayed the bands of mPEG combined with RBC memberane protein and the slow mobility of membrane protein. The hemagglutination of PEGylation RBC ghosts did not take place and mPEG still covered the antigen. In conclusion, mPEG-SPA can bind the erythrocyte with its extracted membrane protein in both ghosts and living erythrocytes. 　　Key words mPEG modified red blood cell; Rh antigen; red blood cell 　　mPEG-SPA 可以与蛋白质和多肽中的赖氨酸（K）、精氨酸(R)的胺基、组氨酸(H) 的咪唑基以及酪氨酸(Y)的羟基发生化学反应，形成共价结合键，从而结合到蛋白质多肽上［1］，达到遮蔽抗原的作用。mPEG与红细胞膜Rh抗原结合后是否稳定，进入人体后是否会脱落，这些问题关系到修饰后红细胞进入人体后的安全性。为此，我们采用mPEG修饰的红细胞膜蛋白SDS电泳、红细胞血影凝集实验以及用4℃ CPD保养