脾内免疫制备抗人血小板第4因子单克隆抗体和鉴定

Preparation and Identification of a Monoclonal Antibody against Human Platelet Factor 4 by Intrasplenic Immunization

目的制备抗人血小板第４因子单克隆抗体用于ＰＦ４的基础研究和临床血小板活化状态的评价。方法用常规和脾内免疫相结合的方法免疫Ｂａｂｌ／ｃ小鼠，经细胞融合和ＥＬＩＳＡ筛选后，进行单抗的特性和特异性研究。结果成功制备了一株抗人 ＰＦ４单克隆抗体（ＳＺ－ ９５），ＥＬＩＳＡ显示上清和腹水效价分别为 ２ × １０-3和 ５ × １０-6，Ｗｅｓｔｅｍｂｌｏｔ证实ＳＺ－９５特异性识别人血小板第４因子。结论常规免疫结合脾内免疫是制备抗低分子量保守蛋白质单抗的有效方法，制备的ＳＺ－９５具有科研运用和临床应用的良好前景。

Objective To prepare monoclonal antibodies against PF4 used in research work and clinical diagnosis. Methods Babl/c mouse was immunized by intraperitoneal and intrasplenic routes. After hybridizing and screening,we charactered MoAbs by ELISA and western blot. Results A MoAb (SZ - 95) against PF4 hybridizing was obtained. The biers of this MoAn in culture supernatants and ascites fluid were 2 × 10-3 and 5 ×10-6 respectively by indirect ELISA. Western blot assay demonstrated SZ - 95 recognized PF4 efficiently under reduc condition. Conclusion Using intrasplenic after intraperitioneal route is an efficient method to prepare MoAbs against a low molecular weight and highly conserVed protein such as PF4. SZ - 95 can serve as a beneficial tool for the study of PF4 and its clinical application.