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目的：制备共刺激分子ＣＤ８０单抗并鉴定其生物学活性。方法；采用给经ＭＤＡ４５３细胞免疫的小鼠注射环磷酰胺１００ｍｇ／（ｋｇ·次）诱导对ＭＤＡ４５３细胞的免疫耐受基础上，再用已建立能稳定表达ｈＣＤ８０分子的ＭＤＡ４５３细胞免疫小鼠，取经检测抗体阳性强的小鼠脾细胞，按常规方法制备单抗。通过Ｔ增殖阻断实验鉴定其活性。结果：ＣＥＬＬ－ＥＬＩＳＡ、ＦＡＣＳ鉴定获得一株抗ＣＤ８０单抗，腹水效价为１０－６，属ＩｇＧ１亚类。Ｔ细胞增殖阻断试验显示，此单抗能部分阻断Ｔ细胞对相应刺激的增殖反应。结论：应用环磷酰胺免疫删减法成功制备膜抗原ＣＤ８０单抗，该单抗有一定应用前景。
Objective: To prepare costimulatory molecule CD80 monoclonal antibody and identify its biological activity. Methods: The mice immunized with MDA453 were injected with cyclophosphamide 100mg / (kg · times) on MDA453 cells on the basis of the immune tolerance, and then established a stable expression of hCD80 molecules MDA453 cells were immunized mice Detection of antibody-positive mouse spleen cells, monoclonal antibodies prepared by conventional methods. Its activity was identified by T proliferation blocking assay. Results: CELL-ELISA, FACS identified an anti-CD80 monoclonal antibody, ascites titer of 10-6, is a subclass of IgG1. T cell proliferation block test showed that this monoclonal antibody partially blocked the T cell proliferation response to the corresponding stimulus. Conclusion: CD80 mAb is successfully prepared by immunospot subtraction of cyclophosphamide. The monoclonal antibody has certain application prospects.