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目的建立三氯醋酸(Trichloroacetic acid,TCA)沉淀法测定食蟹猴体内125I-rhIL-12的含量,并进行验证及初步应用。方法采用Na125I标记结合TCA沉淀蛋白的方法测定食蟹猴体内125I-rhIL-12的含量,对建立的方法进行线性、精密度及回收率验证;将2μg/kg125I-rhIL-12单剂量皮下注射食蟹猴,分别于注射后4、8、24、32、48、72、96、120、144和168 h收集尿和粪,用建立的方法测定放射性,计算排出放射性占注入放射性的百分比。结果125I-rhIL-12的效价与未标记对照品效价无显著差异;在0.02～200 ng-当量范围内,不同组织/体液中125I-rhIL-12的加入量与总放射性及TCA沉淀放射性之间线性关系良好,变异系数(CV)多小于20%;125I-rhIL-12原形主要在TCA沉淀部分。125I-rhIL-12经皮下注射食蟹猴后,放射性主要经尿排泄,少量经粪排泄,皮下注射后168 h,可排泄完全。结论125I与rhIL-12在体内结合稳定,TCA沉淀法可用于测定食蟹猴体内125I-rhIL-12的含量。
Objective To establish a method for the determination of 125I-rhIL-12 in cynomolgus monkeys by trichloroacetic acid (TCA) precipitation method, and verify and preliminary application. Methods The content of 125I-rhIL-12 in cynomolgus monkeys was determined by Na125I-labeled and TCA-precipitated protein. The linearity, precision and recovery of the established method were validated. A single dose of 2μg / kg125I-rhIL-12 Urine and faeces were collected at 4, 8, 24, 32, 48, 72, 96, 120, 144 and 168 h after injection respectively. The radioactivity was measured by the established method and the percentage of radioactivity discharged was calculated. Results The titer of 125I-rhIL-12 was not significantly different from that of untreated control. The addition of 125I-rhIL-12 in different tissues / body fluids in the range of 0.02 ~ 200 ng- (CV) was less than 20%. The original form of 125I-rhIL-12 was mainly in the precipitation part of TCA. 125I-rhIL-12 subcutaneously injected cynomolgus monkeys, the main radioactive excretion of urine, a small amount of manure excretion, 168 h after subcutaneous injection, can be excreted completely. Conclusion 125I and rhIL-12 are stable in vivo. The TCA precipitation method can be used to determine the content of 125I-rhIL-12 in cynomolgus monkeys.