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目的:制备抗心型脂肪酸结合蛋白(H-FABP)单克隆抗体(mAb),并建立侧向免疫层析方法检测血浆中H-FABP。方法:用H-FABP蛋白免疫纯系Balb/c小鼠,采用杂交瘤技术建立能稳定分泌抗人H-FABP的单克隆杂交瘤细胞株。常规制备腹水,纯化后得到特异性抗H-FABP单克隆抗体,进行效价、特异性、亲和力的鉴定分析,并在ELISA平台进行抗体配对,用所筛选到的抗体对初步建立了检测H-FABP的侧向免疫层析方法。结果:成功获得12株稳定分泌抗体的阳性细胞株,并筛选出能相互配对,并应用于侧向免疫层析平台的抗体3D1和5F4,检测临床样品与对照试剂比较总符合率为100%。结论:筛选能稳定分泌抗体的细胞株,配对抗体应用于侧向免疫层析检测方法中,能快速、特异、灵敏的检测出临床样品中H-FABP,为临床应用快速检测H-FABP指标提供了方法和关键材料。
OBJECTIVE: To prepare monoclonal antibody (mAb) against heart-type fatty acid binding protein (H-FABP) and to establish a lateral immunochromatographic assay for detecting H-FABP in plasma. Methods: The pure Balb / c mice were immunized with H-FABP protein, and the monoclonal hybridoma cell line stably secreting anti-human H-FABP was established by hybridoma technique. The ascites was prepared routinely and the specific anti-H-FABP monoclonal antibody was obtained after purification. The titer, specificity and affinity of the monoclonal antibody were identified and analyzed by ELISA. The antibody against H- FABP lateral immunochromatography. RESULTS: Twelve positive cell lines stably secreting antibodies were successfully obtained and 3D1 and 5F4 antibodies that could be paired with each other and applied to the lateral immunochromatographic platform were screened out. The total coincidence rate of clinical samples and control reagents was 100%. Conclusion: Screening of cell lines that can stably secrete antibodies, the paired antibodies used in lateral immunochromatography detection method, rapid, specific and sensitive detection of H-FABP in clinical samples for the rapid detection of H-FABP indicators for clinical applications The method and the key material.