Ruan K, Hashida S, Yoshitake S, Ishikawa E, Wakisaka O, Yamamoto Y, Ichioka T, Nakajima K
Ann Clin Biochem. 1986 Jan;23 ( Pt 1):54-8. doi: 10.1177/000456328602300106.
This paper describes an improved sandwich enzyme immunoassay for insulin in human serum. The detection limit was significantly improved from 0.1 mU/L to 0.02 mU/L, by incubation with guinea-pig anti-insulin Fab'-peroxidase conjugate in the presence of normal guinea-pig F(ab')2 to reduce the non-specific binding of the conjugate. The degree of serum interference was considerably reduced by coating polystyrene balls with acid-treated anti-insulin IgG and by incubating the polystyrene balls with serum samples at a lower temperature for a shorter time. The time for incubation with anti-insulin Fab'-peroxidase conjugate was also shortened. As a result, the volume of serum that could be used increased from 10 microliters to 50 microliters, and the time for immuno-reactions was reduced from 8 h to 4 h. Applicability of the present improvement to enzyme immunoassay of other antigens in human serum is discussed.
本文描述了一种用于检测人血清中胰岛素的改进型夹心酶免疫测定法。通过在正常豚鼠F(ab')2存在下与豚鼠抗胰岛素Fab'-过氧化物酶缀合物孵育，以减少缀合物的非特异性结合，检测限从0.1 mU/L显著提高到0.02 mU/L。通过用酸处理的抗胰岛素IgG包被聚苯乙烯球，并在较低温度下较短时间内将聚苯乙烯球与血清样品孵育，血清干扰程度大大降低。与抗胰岛素Fab'-过氧化物酶缀合物的孵育时间也缩短了。结果，可使用的血清体积从10微升增加到50微升，免疫反应时间从8小时减少到4小时。讨论了本改进方法在人血清中其他抗原酶免疫测定中的适用性。
