Molafilabi Azam, Shahabi Majid, Rafatpanah Houshang, Mashkani Baratali
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, IBTO Bldg, Hemmat Exp. Way, 1449613111 Tehran, Iran.
2Immunology Research Center, Inflammation and Inflammatory Diseases Division, Mashhad University of Medical Sciences, 9177948564 Mashhad, Iran.
Indian J Hematol Blood Transfus. 2019 Jan;35(1):125-130. doi: 10.1007/s12288-018-0999-9. Epub 2018 Aug 11.
Enzymatic removal of blood groups antigens A and B is an efficient method for production of universal red blood cells. In this research, an α--acetylgalactosaminidase (NAGA) enzyme was expressed in for digestion of the A blood antigen. DNA sequence of the gene NAGA, originally expressed in Elizabethkingia meningosepticum (NAGA-EM), was ordered for optimization and synthesis. It was then expressed in (KM71H and GS115 strains). Expression of the recombinant NAGA was evaluated by dot blot, SDS-PAGE, and Western blotting. The activity of the enzyme was measured using a synthetic substrate in addition to the conversion of group A red blood cells to the O cells. Expression of NAGA-EM with an apparent molecular mass of 55 kDa was verified by dot blot, SDS-PAGE and Western blot analysis. The maximum enzyme activity in the supernatant of KM71H was higher than that in the GS115 (250 vs. 200 U/ml). Treated group A RBCs did not react with the anti-A antiserum or with the sera from individuals with blood groups B and O. The results of this study indicated that NAGA-EM is an efficient enzyme for production of universal O blood cells.
酶法去除血型抗原A和B是生产通用型红细胞的有效方法。在本研究中，一种α-乙酰半乳糖胺酶（NAGA）在[具体宿主菌名称未给出]中表达以消化A血型抗原。最初在脑膜伊丽莎白菌（NAGA-EM）中表达的NAGA基因的DNA序列被订购用于优化和合成。然后在[具体宿主菌名称未给出]（KM71H和GS115菌株）中表达。通过斑点印迹、SDS-PAGE和蛋白质免疫印迹评估重组NAGA的表达。除了将A型红细胞转化为O型细胞外，还使用合成底物测量该酶的活性。通过斑点印迹、SDS-PAGE和蛋白质免疫印迹分析验证了表观分子量为55 kDa的NAGA-EM的表达。KM71H上清液中的最大酶活性高于GS115中的最大酶活性（250对200 U/ml）。处理后的A型红细胞不与抗A抗血清或B型和O型血个体的血清发生反应。本研究结果表明，NAGA-EM是生产通用型O型血细胞的有效酶。
