酶联免疫吸附测定用于胰岛素生产效价检测

ELISA OF INSULIN IN MANUFACTURING PROCESS

用高碘酸钠氧化法将辣根过氧化物酶标记于胰岛素。用亲和层析法纯化豚鼠抗猪胰岛素抗体。以ELISA-抗原竞争法测定猪胰岛素生产过程中间品效价。胰脏中非胰岛素成分对测定无干扰。对于生物效价的符合程度与RIA相当。在50-800μIU/ml测量范围内,测量偏差为10-20％。还讨论了对测定有影响的若干因素,如抗体包被时间、孵育时间、酶反应时间及非特异性吸附等。

Insulin was labeled with horseradish peroxidase by sodium periodate oxidation method. Guinea pig antibody against pig insulin was purified by affinity chroma- tography. Quantitative detection of insulin in different stages of manufacturing was carried out with ELISA (Enzyme linked Immunosorbent Assay) using antige- nenzyme conjugate. The components other than insulin in the pancreas did not interfere the result in ELISA. When the biological effective units detected by ELISA were in a range of 50-800 μIU/ml, the deviations were about 10-20%. All the influenced factors, such as the time of antibody adsorption, incuba- tion, reaction of enzyme and non-specific adsorption were discussed.