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目的 :探讨NBS法标记抗CD2 0F(ab’) 2 的最优条件。方法 :其它条件固定时 ,依次改变反应时间、NBS用量、反应温度、反应体积 ,进行标记后将反应混合物经PD - 1 0柱淋洗 ,收集蛋白峰 ,测定放射性计数 ,计算标记率和免疫活性分数。结果 :反应时间在 2min之前 ,标记率随时间延长而提高 ,2min后出现平台 ,3min达最大值 ,活性在 3min之前变化不大 ,之后明显降低 ;NBS/F(ab’) 2 在 1 6时标记率最高 ,1 4之后活性急剧下降 ;温度对标记率和活性的影响不明显 ;小体积反应有利于提高标记率。结论 :反应时间为 3min ,NBS/F(ab’) 2 为1 6 ,室温小体积标记为最优条件
Objective: To investigate the optimal conditions of labeling anti-CD20F (ab ’) 2 by NBS method. Methods: When the other conditions were fixed, the reaction mixture was sequentially rinsed with PD - 10 column after changing the reaction time, the amount of NBS, the reaction temperature and the reaction volume. The peak of the protein was collected and the radioactivity count, the labeling rate and the immunocompetence fraction. Results: The reaction time was up to 2min and the labeling efficiency increased with time. After 2min, the plateau appeared and reached the maximum at 3min. The activity did not change much before 3min and then decreased significantly. The NBS / F (ab ’ The labeling rate was the highest, and the activity dropped sharply after 14 days. The effect of temperature on labeling rate and activity was not obvious. The small volume reaction was helpful to improve the labeling rate. Conclusion: The reaction time is 3 min, the NBS / F (ab ’) 2 is 16, and the small volume at room temperature is labeled as the optimal condition