LAIR-2单克隆抗体的制备、鉴定和夹心ELISA方法的建立

为了制备LAIR 2单克隆抗体并建立夹心ELISA方法。应用淋巴细胞杂交瘤技术制备抗LAIR 2单克隆抗体 (mAb )。采用间接免疫荧光染色和流式细胞术鉴定LAIR 2的细胞分布。辣根过氧化物酶 (HRP )标记LAIR 2mAb ,并建立检测LAIR 2的夹心ELISA方法。用重导向杀伤实验 (RCA )鉴定LAIR分子参与调节杀伤功能的关系。结果成功地制备了 3株特异识别LAIR 2的mAb ,1株mAb (3G4 )能够识别LAIR 1和LAIR 2的共同表位 ,但不能在重导向杀伤实验中抑制CD16诱导的杀伤功能。夹心ELISA方法检测LAIR 2的敏感性为 5ng/ml,为LAIR 2的分布及定量检测提供了方法

Preparation and Identification of Monoclonal Antibodies Against Leukocyte-Associated Immunoglobulin-Like Receptor-2 (LAIR-2) and Establishment of Sandwich ELISA for Detecting LAIR-2

To prepare monoclonal antibodies(mAb) against LAIR-2 and establish sandwich ELISA to detect LAIR-2,distribution of LAIR-2 was investigated with indirect fluorescence staining and flow cytomytry analysis. Based on epitope mapping of LAIR-2 and LAIR-2 mAb HRP-conjugation, the sandwich ELISA detecting LAIR-2 was established. The potential role played by LAIR-2 mAb 3G4 in cytotoxicity function was identified with redirected cytotoxicity assay(RCA). Four strains of mAb recognizing LAIR-2-myc fusion protein were obtained by routine hybridoma technique. Among the four strains of mAb, one clone named 3G4 can recognize both LAIR-1 and LAIR-2, but it couldn't inhibit the cytotoxicity induced by CD16 mAb in RCA. The limitation measured by sandwich ELISA for detecting LAIR-2 reached to 5 ng/ml, which may be useful for basic and clinical research on LAIR-2 distribution and quantitative analysis.