马来酰亚胺苯甲胺酯聚乙二醇遮盖红细胞RhD抗原研究

STUDY ON THE CHEMICAL CAMOUFLAGE OF RhD ANTIGENS OF ERYTHROCYTES WITH MALEIMIDOPHENYL POLYETHYLENE GLYCOL

[目的]马来酰亚胺苯甲胺酯聚乙二醇(MDPEG)修饰红细胞,遮蔽RhD抗原,使Rh+红细胞转变为Rh-红细胞。[方法]甲基苯甲酸和聚乙二醇单甲醚(分子量5000)为原料合成MDPEG(分子量5910)。红细胞修饰:3%～5%RBC+MDPEG+2-亚氨基硫烷(IT)、pH 7.4、室温2 h。盐水法进行红细胞与ABO血型、RhD血型血清学实验。血流变仪测定红细胞黏度等指数。盐水梯度法测定红细胞渗透脆性。[结果]30 mg/ml马来酰亚胺苯甲胺酯聚乙二醇能有效遮蔽3%～5%红细胞RhD抗原,红细胞与抗RhD抗体无凝集反应,光学显微镜观察红细胞无聚集,细胞形态正常。红细胞膜渗透性、红细胞聚集指数轻微升高,刚性指数轻微下降。[结论]马来酰亚胺苯甲胺酯聚乙二醇(MD-PEG)能完全遮盖RhD抗原,获得RhD阴性红细胞,细胞形态,结构正常。

[Objective]To mask RhD antigens of RBCs by RBC-PEGylation with maleimidophenyl-polyethylene glycol（MDPEG）,and to convert Rh＋ RBCs into Rh-RBCs.[Methods]MDPEG（Mw 5910）was synthesized from 4-aminobenzoic acid and methylether polyethylene（Mw 5 000）.RBC-PEGylation：3%-5% RBC＋MDPEG＋2-Iminothiolane（IT）with pH of 7.4,at room tempterture for 2 hours.RBCs were reacted with ABO and RhD antibodies in normal saline,the viscosity of RBCs was tested with apparatus.The erythrocyte osmotic fragility was tested in different saline concentrations.[Results]30mg/ml MDPEG was enough to mask 3%-5% of the RhD antigens of RBCs,MDPEG-RBCs were negative with anti-RhD antibodies,and with no agglutination under light microscope.RBCs were morphologically normal.The erythrocyte osmotic fragility and aggregation index slightly increased,but its rigid index slightly decreased.[Conclusion]RhD antigens of RBCs can be masked completely by MDPEG to get RhD negative RBCs.The infrastructure and morphology of the RhD negative RBCs are normal.