Nakajima Y, Matsuno S, Noto N, Saitoh Y, Sato T
Tohoku J Exp Med. 1980 Jun;131(2):119-26. doi: 10.1620/tjem.131.119.
An improved method of radioimmunoassay was devised to offer a successful formula for determining blood concentration of elastase. With porcine pancreatic elastase as the antigen, rabbits were immunized to obtain antiserum. Iodinated elastase labeled by the chloramine-T procedure using 131I (or 125I) had a specific activity of 200-300 mCi/mg. The double antibody method was used for BF separation. While the usual method of radioimmunoassay was not always successful in obtaining accurate serum concentration of elastase, the use of diisopropyl fluorophosphate (DFP) was able to eliminate the disturbing influence of intra-serious inhibitors, alpha 1-AT and alpha 2-M, eventually producing satisfactory results. With the use of DFP, the elastase standard curve and the porcine serum dilution curve had a statistically significant correlation; precision and recovery were both satisfactory; cross-reactivity of the antiserum with trypsin and chymotrypsin was less than 0.001%. The minimal detectable concentration of elastase was 5 ng/ml, and the range of normal fasting porcine serum level was 70-100 ng/ml.
设计了一种改进的放射免疫测定方法，为测定弹性蛋白酶的血药浓度提供了一个成功的方案。以猪胰弹性蛋白酶为抗原，免疫家兔获得抗血清。用131I（或125I）通过氯胺-T法标记的碘化弹性蛋白酶的比活性为200-300mCi/mg。采用双抗体法进行BF分离。虽然常规的放射免疫测定方法在获得准确的血清弹性蛋白酶浓度方面并不总是成功的，但使用二异丙基氟磷酸酯（DFP）能够消除血清内抑制剂α1-AT和α2-M的干扰影响，最终产生令人满意的结果。使用DFP时，弹性蛋白酶标准曲线与猪血清稀释曲线具有统计学显著相关性；精密度和回收率均令人满意；抗血清与胰蛋白酶和糜蛋白酶的交叉反应率小于0.001%。弹性蛋白酶的最低检测浓度为5ng/ml，正常空腹猪血清水平范围为70-100ng/ml。
