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选用正交设计 L_(16)(4~5)表对影响~(125)I 标记 SOD 的5个因素(标记温度、氯胺T 加入量、标记反应时间、PB 缓冲液浓度和缓冲液体积),每个因素的4个不同水平进行了研究。标记产物~(125)I-SOD 用 Sephadex G25色层柱进行分离,测定~(125)I 的标记率和~(125)I-SOD的相对活度,仅做16组试验,即可确定最佳标记条件。试验结果表明,对50μgSOD,标记温度为10℃,氯胺 T 加入量为30μg,标记反应时间为2min,PB 溶液浓度为0.1mol/L,PB 溶液体积为50μl 时,~(125)I 的标记率可达75.9%,~(125)I-SOD 的相对活度可保持在94.7%;用纸色层法分析了~(125)I-SOD 产品的放射化学纯度,可达95%以上;估算了~(125)I-SOD 产品的比活度,为～1.12×10~5Bq/μg SOD。
Five factors (labeling temperature, addition of chloramine T, labeling reaction time, concentration of PB buffer and volume of buffer) affecting 125 I-labeled SOD were studied by orthogonal design L_ (16) Four different levels of each factor were studied. The labeled product 125 I-SOD was separated on a Sephadex G25 chromatographic column to determine the labeling rate of ~ (125) I and the relative activity of ~ (125) I-SOD. Good mark conditions. The results showed that the labeling of ~ (125) I for 50μgSOD, labeling temperature 10 ℃, chloramine T dosage 30μg, labeling reaction time 2min, PB solution concentration 0.1mol / L, PB solution volume 50μl The relative activity of ~ (125) I-SOD was maintained at 94.7%. The radiochemical purity of ~ (125) I-SOD product was over 95% The specific activity of ~ (125) I-SOD product was ~ 1.12 × 10 ~ 5Bq / μg SOD.