单克隆抗M、抗N抗体的研制及初步应用

用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。

PREPARATION AND APPLICATION OF MONOCLONAL ANTI-M AND ANTI-N ANTIBODIES

BALB/c mice were immunized by intraperitoneal injections with N-grouperythroeytes washed five times with saline and purified glycophorin A.Immu-nized spleen cells were fused with SP 2/0 murine myoloma cells.After 10-15days of incubation at 37℃,the medium from each well was examined for anti-bodies by hemagglutination reaction by using OM and ON RBC.The hybridomacells secreting high titer and specific monoclonal anti-M and anti-N antibodieswere established.These hybridoma cells have the ability of constant secretingantibodies which belong to the IgG class,and specifically reacted with eitherhuman group N erythrocytes or human group M erythrocytes and old blood stainsamples.Monoclonal anti-M and anti-N sera gave identical results with comm-ercial immunized anti-M and anti-N tying sera from rabbits.