B→O血型转换批量制备通用型红细胞的研究

目的建立B→O血型转换红细胞(通用型红细胞)批量制备工艺,为临床提供通用型红细胞制品。方法用不同pH值的缓冲液对1个使用单位(200ml)的B型红细胞进行预处理。在专门设计的转型密闭处理系统中,用α半乳糖苷酶对B型红细胞表面B抗原进行酶解,并检测酶解后红细胞ABO血型抗原及结构、功能。结果酶解前用pH4.2缓冲液预处理组上清游离Hb含量显著低于pH2.8缓冲液处理组;经α半乳糖苷酶(100Uml红细胞)在pH5.6,26℃条件下酶解60min,红细胞B型抗原被完全清除,转换为O型;红细胞形态、变形性、ATP、2,3DPG、胆固醇含量、乙酰胆酐酯酶活性、渗透脆性、pH、上清游离Hb、K+、Na+含量与对照组(正常红细胞)比较差异无显著意义,均在正常范围。结论建立了B→O血型转换通用型红细胞的批量制备工艺。通用型红细胞结构、功能、形态均正常,细菌、热原实验合格。

Batch Preparation of Universal Red Blood Cells Converted from Group B to Croup O

Objective To develop a procedure for batch preparation of universal red blood cells converted from group B to group O and provide universal red blood cells for clinic.Methods Pre-treat a full unit(200 ml) of red blood cell group B with the buffers at different pH values.In a specially designed enclosed system,digest the antigen B on the surface of human red blood cell group B with recombinant galactosidase-α and subject the digested red blood cells to ABO grouping as well as studies on structure and function.Results The Hb content in supernatant of red blood cells pre-treated with buffer(pH 4.2) was significantly lower than that pre-treated with buffer(pH 2.8).After digestion with galactosidase-α(100 U/ml) at pH 5.6,26℃ for 60 minutes,the antigen B was removed completely,so the red blood cells were converted from group B to group O.The quality indexes of converted red blood cells,such as morphology,deformability,the contents of ATP,2,3-DPG and cholesterol,acetylcholinesterase activity,osmotic fragility,pH as well as the contents of free Hb,free potassium ion and free sodium ion in supernatant,were within a normal range,and showed no significant difference from those of normal red blood cells.Conclusion A procedure for batch preparation of universal red blood cells converted from group B to group O was successfully developed.The prepared universal red blood cells were normal in structure,function and morphology and qualified in bacterium and pyrogen tests.