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目的：建立人抗凝血酶Ⅲ（ Ａ Ｔ Ⅲ）的 Ｅ Ｌ Ｉ Ｓ Ａ 检测试剂盒及 Ａ ＴⅢ的纯化提供试剂准备。方法：用 Ａ ＴⅢ进行 ２ 次基础免疫、２ 次加强免疫的方法免疫 Ｂａｌｂ／ｃ 小鼠，经间接 Ｅ Ｌ Ｉ Ｓ Ａ 的方法筛选杂交瘤细胞株，通过吸收试验、免疫印迹试验等进行特异性分析。结果：筛选出１５ 株分泌抗人 Ａ ＴⅢ单克隆抗体的杂交瘤细胞株，对其中３ 株（ Ｂ４、 Ｂ５、 Ｂ７）作进一步研究，３ 株杂交瘤细胞分泌抗体亚型均为 Ｉｇ Ｇ１ 型，３ 株细胞培养上清滴度平均达到４×１０３，小鼠腹水滴度平均达到 ４×１０６，３ 株抗体均具抗人 Ａ ＴⅢ的特异性。结论：所筛选的单抗为 Ｅ Ｌ Ｉ Ｓ Ａ 检测试剂盒的制备及 Ａ ＴⅢ的纯化提供了试剂准备。
OBJECTIVE: To establish a human antithrombin Ⅲ (A T Ⅲ) E L I S A detection kit and A T Ⅲ purification to provide reagent preparation. Methods: Balb / c mice were immunized twice with basic immunization with A T Ⅲ and twice with booster immunization. The hybridoma cell lines were screened by indirect ELISAs. The specificities of the hybridomas were determined by absorption test and western blotting analysis. Results: Fifteen hybridoma cell lines secreting anti-human A TⅢ monoclonal antibodies were screened out and 3 of them (B4, B5 and B7) were further studied. The antibody subtypes secreted by all three hybridomas were Ig G1 type, The titers of the three cell culture supernatants reached an average of 4 × 103, the ascites titers of mice reached an average of 4 × 106, and the three antibodies showed anti-human A TⅢ specificity. Conclusion: The McAbs screened for the preparation of ELISA kit and the preparation of A T Ⅲ.