生物素化抗细胞因子单克隆抗体的制备

目的 探讨生物素化单克隆抗体的制备方法。方法 应用经超滤、硫酸铵沉淀、ProteinG亲和层析纯化的来源于杂交瘤细胞培养上清中的大鼠抗小鼠IL-5、IL-4和IFN-γ单克隆抗体，以每mg抗体加入生物素制剂80ug室温反应60min，其反应液经SephadexG-25层析纯化，然后用生物素-亲和素ELISA法测定所制备的生物素化抗体活性。结果 该法制备的生物素化抗体活性高，用于检测感染巴西钩虫的IL-5转基因小鼠和非转基因小鼠的血清标本敏感性好。结论 该生物素化抗体的抗备方法具有制备简单、稳定性好等优点。

Preparation of biotinylated anti-cytokine monoclonal antibodies

Objective To explore a method of preparation for biotinylatedmonoclonal antibodies． Methods Rat anti-mouse IL-5， IL-4 and IFN-γ were purified by ultra filtration， precipitated with saturated ammonium sulfate further purified by chromatography with the protein G-sepharose affinity column from the hybridoma cell culture supernatants． We added 80 μg of the biotinylation reagent per 1 mg of the monoclonal antibody， and incubated them at room temperature for 60 min． After that， we applied the monoclonal antibody sample to the Sephadex G-25 column and got the purified biotinylated monoclonal antibody． The antibodies were tested the activities through the Avidin-biotin ELISA． Results The results showed that the antibodies kept high biological activities and sensitivities in detecting the sera from IL-5 transgenic mice and non-transgenic mice by ELISA． Conclusion The method is simple and stable．