Enander I, Matsson P, Nystrand J, Andersson A S, Eklund E, Bradford T R, Schwartz L B
Research Department, Pharmacia Diagnostics AB, Uppsala, Sweden.
J Immunol Methods. 1991 Apr 8;138(1):39-46. doi: 10.1016/0022-1759(91)90062-k.
A solid phase immunoradiometric assay was developed for the quantitation of tryptase released from activated human mast cells. Tryptase exhibits a linear dose-response curve over the standard range of 2-50 micrograms/l in buffer, serum, and plasma. The dose-response curve approached a plateau at a tryptase concentration of 100 micrograms/l and exhibited partial inhibition at concentrations above 10,000 micrograms/l. The sensitivity of the assay was 0.2-0.4 micrograms/l, and the intra-assay and interassay coefficients of variation were below 4% at 2 micrograms/l or higher tryptase concentrations. The recovery of known amounts of purified tryptase added to serum ranged from 91 to 115%. Detection of tryptase was evaluated with several body fluids and was accurate in sera, plasma, bronchoalveolar lavage fluid, nasal lavage fluid, and saliva. The concentration of tryptase was examined in serum samples from 100 healthy controls; in each case the level was less than 2 micrograms/l. The immunoassay also was utilized to examine serum levels of tryptase after the onset of a hypotensive reaction in one patient receiving general anesthesia. A maximally elevated level of tryptase (25 micrograms/l) was detected at the first time point, 0.5 h, and elevated levels persisted to 6 h before a return to normal levels was documented at 24 h. Thus, the involvement of mast cell activation in hypotensive subjects can be ascertained by this new tryptase radioimmunoassay.
已开发出一种固相免疫放射分析方法，用于定量检测活化的人肥大细胞释放的类胰蛋白酶。在缓冲液、血清和血浆中，类胰蛋白酶在2-50微克/升的标准范围内呈现线性剂量反应曲线。在类胰蛋白酶浓度为100微克/升时，剂量反应曲线接近平稳期，在浓度高于10,000微克/升时表现出部分抑制。该分析方法的灵敏度为0.2-0.4微克/升，在类胰蛋白酶浓度为2微克/升或更高时，批内和批间变异系数均低于4%。添加到血清中的已知量纯化类胰蛋白酶的回收率在91%至115%之间。用几种体液评估类胰蛋白酶的检测情况，在血清、血浆、支气管肺泡灌洗液、鼻腔灌洗液和唾液中检测结果准确。检测了100名健康对照者血清样本中的类胰蛋白酶浓度；在每种情况下，水平均低于2微克/升。该免疫分析方法还用于检测一名接受全身麻醉的患者发生低血压反应后血清中的类胰蛋白酶水平。在第一个时间点0.5小时检测到类胰蛋白酶的最大升高水平（25微克/升），升高水平持续到6小时，之后在24小时记录到恢复正常水平。因此，通过这种新的类胰蛋白酶放射免疫分析方法可以确定低血压患者中肥大细胞活化的情况。
