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目的：制备抗人红细胞膜血型糖蛋白Ａ（ＧＰＡ）的特异单克隆抗体，对研究ＧＰＡ基因突变和ＧＰＡ蛋白糖基化变异有重要意义。方法：分别用ＯＭ和纯化Ｍ－ＧＰＡ蛋白以及ＯＮ型红细胞和纯化Ｎ－ＧＰＡ蛋白免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与小鼠骨髓瘤细胞Ｓｐ２／０融合，经正常和酶处理人细胞凝集筛选，间接免疫荧光，酶联免疫分析及蛋白质印迹鉴定，制备了鼠抗人ＧＰＡ的单克隆抗体。结果：获得抗ＧＰＡ蛋白１～３９位肽段及相关糖链的５株单抗，除２Ｅ９以外均为糖链依赖型。其中抗Ｍ，Ｎ－ＧＰＡ单抗２株：２Ｅ９Ｉｇ（Ｇ１，λ）和９Ｅ９Ｉｇ（Ｇ３，κ）；２Ｅ９抗Ｍ，Ｎ－ＧＰＡ６－３９位同源肽段，９Ｅ９抗Ｍ，Ｎ－ＧＰＡ１～２６位肽段。Ｎ－ＧＰＡ单抗３株：３Ｃ３Ｉｇ（Ｇ１，κ），５Ｆ９Ｉｇ（Ｇ３，κ）和６Ａ８Ｉｇ（Ｇ３，λ），均为抗Ｎ－ＧＰＡ１～６位肽段抗体。４株单抗与胰蛋白酶处理的红细胞不能反应，６Ａ８使胰蛋白酶处理的Ｍ型红细胞凝集而不能凝集胰蛋白酶处理的Ｎ型红细胞，推测Ｍ和Ｎ型红细胞的ＧＰＢ蛋白存在某种差异。结论：这些抗体对研究ＧＰＡ基因突变和蛋白糖基化变异有应用价值
OBJECTIVE: To prepare specific monoclonal antibodies against human erythrocyte glycoprotein A (GPA), which is of great significance for the study of GPA mutation and glycosylation variation of GPA. METHODS: BALB / c mice were immunized with OM and purified M-GPA protein and purified erythrocyte and purified N-GPA protein respectively. Sp2 / 0 cells were fused with mouse myeloma cells. Normal and enzyme-treated human cells were aggregated Screening, indirect immunofluorescence, enzyme-linked immunosorbent assay and Western blotting, the monoclonal anti-human GPA antibody was prepared. Results: The five monoclonal antibodies against GPA peptides 1 to 39 and their related sugar chains were all sugar chain-dependent except for 2E9. Among them, 2 strains of anti-M and N-GPA McAbs: 2E9Ig (G1, λ) and 9E9Ig (G3, κ); 2E9 anti M and N-GPA 6-39 homologous peptides; 9E9 anti-M and N-GPA1-26 Bit peptide. N-GPA McAb 3 strains: 3C3Ig (G1, κ), 5F9Ig (G3, κ) and 6A8Ig (G3, λ), were anti-N-GPA1 ~ 6 peptide antibody. Four monoclonal antibodies did not react with trypsin-treated erythrocytes. 6A8 aggregated trypsin-treated M-type erythrocytes to trypsin-free N-type erythrocytes and speculated that there was a difference in GPB protein between M and N-type erythrocytes. Conclusions: These antibodies have utility in the study of GPA mutations and glycosylation variants