Solid-phase radioimmunoassay for human neutrophil elastase: a sensitive method for determining secreted and cell-associated enzyme.
Solid-phase radioimmunoassay for human neutrophil elastase: a sensitive method for determining secreted and cell-associated enzyme.
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DOI：
10.1016/0003-2697(85)90436-1
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发表时间：
1985-10
影响因子：
2.9
通讯作者：
T. Dunn;W. Blackburn;W. Koopman;L. Heck
中科院分区：
文献类型：
--
作者：
T. Dunn;W. Blackburn;W. Koopman;L. Heck
建立了测定中性粒细胞弹性蛋白酶0.08 ~ 4 ng/ml的固相放射免疫法。通过反复免疫山羊，产生了一种能抑制弹性蛋白溶解的单特异性沉淀抗体。然后获得该血清的IgG部分和亲和纯化抗体，并用于开发该放射免疫测定。放射性标记抗血清与乳铁蛋白、组织蛋白酶G或氨基酸末端同源的丝氨酸蛋白酶结合无交叉反应性。虽然与二异丙基氟磷酸盐处理的中性粒细胞弹性酶相比，血清会影响催化活性中性粒细胞弹性酶的测量，但抗原弹性酶仍可在体液中测量。此外，该试验比市售的用于定量中性粒细胞弹性酶功能活性的底物更敏感。我们发现这种定量分析在平衡研究中非常有用，可以测量分泌的和细胞相关的弹性酶，也可以筛选生物液体中酶的存在。
A solid-phase radioimmunoassay for measuring neutrophil elastase in the range 0.08–4 ng/ml has been developed. A monospecific, precipitating antibody capable of inhibiting elastinolysis was produced by repeated immunizations of a goat. The IgG fraction and affinity-purified antibodies of this serum were then obtained and used to develop this radioimmunoassay. There was no cross-reactivity in binding of the radiolabeled antisera with lactoferrin, cathepsin G, or serine proteinases with amino-terminal amino acid sequence homology. Although serum influences the measurement of catalytically active neutrophil elastase when compared to diisopropylfluorophosphate-treated neutrophil elastase, antigenic elastase may still be measured in body fluids. Furthermore, this assay is more sensitive than commercially available substrates used for quantitating neutrophil elastase by functional activity. We have found this quantitative assay extremely useful in balance studies to measure secreted and cell-associated elastase and in screening of biological fluids for the presence of the enzyme.