温度时间对马来酰亚胺苯甲胺酯聚乙二醇遮蔽Rh抗原的影响

目的观察时间、温度等因素对马来酰亚胺苯甲胺酯聚乙二醇遮盖Rh抗原的影响。方法马来酰亚胺苯甲胺酯聚乙二醇(30mg/ml)、2-亚氨基硫烷盐酸盐(3mg/ml),在pH7.4磷酸钠缓冲液中修饰红细胞(3%-5%),修饰红细胞与ABO/Rh抗体反应,观察马来酰亚胺苯甲胺酯聚乙二醇、乙-亚按基硫烷盐酸盐和红细胞加入的先后顺序以及温度时间对Rh抗原遮盖的效果。结果马来酰亚胺苯甲胺酯聚乙二醇、2-亚氨基硫烷盐酸盐和红细胞加入的先后顺序对红细胞RhD抗原遮盖无影响;但温度、时间对Rh抗原遮盖有明显影响,25-37℃修饰红细胞30min能完全遮蔽Rh抗原,但4℃修饰红细胞120min,仍有部分Rh抗原未被遮盖,显微镜下可见小细胞团块。结论温度、时间对聚乙二醇修饰红细胞遮盖Rh抗原有明显影响,而加入的顺序和间隔时间对抗原修饰无影响。

Influence of time and temperature on Rh antigen masking by maleimidophenyl-polyethylene glycol

Objective To study the effect of temperature and time on the mask of Rh antigen,during the surface decoration of RBC,by maleimidophenyl-polyethylene glycol(MDPEG).Methods RBCs(about 3-5%) were decorated with MDPEG(30mg /ml) and 2-iminothiolane(3mg/ml) in buffer(pH 7.4),and then tested with anti-A,anti-B and anti-RhD.The agglutination of PEG-RBC with anti-RhD was observed with light microscope,and the effect of RhD antigen mask was evaluated.Results The time order of adding MDPEG,2-iminothiolane and RBCs had no significant effect on the Rh antigen mask,while the temperature and time did matter.Decorated at 25℃-37℃ for 30 minutes could completely mask the RhD antigen,while at 4℃ for 120 minutes could only partly mask the RhD antigen,and some small RBC agglutinates could still be observed with microscope.Conclusion MDPEG linking to RBC membrane catalyzed by 2-iminothiolane,to mask RhD antigens on RBCs was a chemical reaction.During the decoration,temperature and time have an important effect on the RhD antigen mask,while the time order of adding MDPEG,2-iminothiolane and RBC does not matter.