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目的:采用国内合成的降钙素基因相关肽(CGRP)及本实验室制备的抗CGRP单克隆抗体建立检测CGRP的放射免疫分析法。方法:酰化试剂法标记CGRP(1-37),采用平衡饱和加样程序法建立放免分析法。结果:纯化后的标记物计算标记率为35％～41％,比活度1000～1200Ci/mmol,放化纯度为93％～98％,免疫活性>92％;该方法的灵敏度为10～20pg/ml,平均批内及批间变异系数分别为11.4％、23.5％。4周后标记物测B_0％为27.5％;初步测32例正常人结果为68.89±15.56pg/ml,范围38.60～112.54pg/ml。结论:本方法基本达到检测要求,但标记物的稳定性及CGRP单抗的稳定性尚需提高。
OBJECTIVE: To establish a radioimmunoassay for detection of CGRP using the domestic synthetic calcitonin gene-related peptide (CGRP) and the monoclonal antibody against CGRP prepared in our laboratory. Methods: CGRP (1-37) was labeled with acylation reagent, and radioimmunoassay was established by the method of equilibrium saturation loading. Results: The calculated labeling rates were 35% ~ 41%, the specific activity was 1000 ~ 1200Ci / mmol, the radiochemical purity was 93% ~ 98% and the immunological activity was> 92%. The sensitivity of this method was 10 ~ 20pg / ml, the average intra-batch and intra-batch coefficient of variation were 11.4%, 23.5%. After 4 weeks, B0% was 27.5%. The initial result of 32 normal subjects was 68.89 ± 15.56pg / ml, ranging from 38.60-112.54pg / ml. Conclusion: The method basically meets the detection requirements, but the stability of the marker and the stability of CGRP monoclonal antibody still need to be improved.