【摘要】 采用双相氯胺-T法对聚乙二醇化重组人白细胞介素-6(PEG-rhIL-6)进行碘化标记,比较了双相氯胺-T法碘化标记PEG-rhIL-6在不同反应条件下的标记效果。用三氯乙酸沉淀法和γ计数探测仪测定标记物的标记率、超滤离心法对标记物进行分离纯化、三氯乙酸沉淀法鉴定标记物的放化纯度、SDS-PAGE电泳分析标记蛋白断链损伤情况。结果表明,采用该方法标记PEG-rhIL-6,在100μL、pH值7.4的反应体系中,控制反应蛋白量为25μg、浓度20g/L的氧化剂15μL、室温反应时间40—50min,可获得较高的标记率和放化纯度,比放射性适中,标记效果比较理想。更多还原

【Abstract】 The two-phase chloramine-T method was applied in the radioiodine-labeling of polyethylene glycol modification of recombinant human interleukin-6 (PEG-rhIL-6). The iodine incorporation was determined by trichloroacetic acid (TCA) and a γ-ray survey meter. The labeled compound was purified by ultrafiltration. The purity of the labeled PEG-rhIL-6 was determined by TCA. The fragmentation of labeled protein was determined by SDS-PAGE-electrophoresis. The labeling efficiencies under different reaction conditions were compared, and the results demonstrated that the optimal conditions for total volume, pH, dosage of added protein, volume of chloramine-T, reaction time and temperature are 100μL, pH=7.4, 25μg, 15μL (20g/L), 40─50 min and room temperature, respectively.更多还原