A SANDWICH ENZYME-IMMUNOASSAY FOR THE DETERMINATION OF NEUTROPHIL LIPOCALIN IN BODY-FLUIDS
A SANDWICH ENZYME-IMMUNOASSAY FOR THE DETERMINATION OF NEUTROPHIL LIPOCALIN IN BODY-FLUIDS
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DOI：
10.1016/0009-8981(95)06020-7
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发表时间：
1995-03-31
影响因子：
5
通讯作者：
TSCHESCHE, H
中科院分区：
文献类型：
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作者：
BLASER, J;TRIEBEL, S;TSCHESCHE, H
从人血沉棕黄层中纯化人中性粒细胞脂质运载蛋白。免疫家兔获得多克隆抗体。抗体与游离脂质运载蛋白以及与PMNL-明胶酶结合的蛋白质反应。该抗体用于建立灵敏的ELISA，用于使用生物素/链霉亲和素系统测定体液中的蛋白质。平均批内C. V.为2.3%，平均批间C. V.为6.7%。人血浆中的回收率测定为98.8%。ELISA允许在0.2-25 μ g/l的浓度范围内测定蛋白质。中性粒细胞脂质运载蛋白浓度的测量结果表明，健康供体的人血浆中含有97 +/- 81 μ g/l（n = 122），血清中的浓度显著更高（P <0.001），为133 +/- 90 μ g/l（n = 122）。在健康供体的尿液中也发现了神经元脂质运载蛋白（8.1 μ g/l; n = 9）。在患有炎性类风湿性关节炎的患者的滑液中发现非常高浓度的这种脂质运载蛋白（1.7 +/-1.4 mg/l; n = 37）。
Human neutrophil lipocalin was purified from human buffycoat. A polyclonal antibody was obtained by immunisation of rabbits. The antibody reacted with the free lipocalin as well as with the PMNL-gelatinase bound protein. This antibody was used to establish a sensitive sandwich-ELISA for the determination of the protein in body fluids using the biotin/streptavidin system. The mean intra-assay C.V. was 2.3% and the mean inter-assay C.V. 6.7%. The recovery in human plasma was determined to be 98.8%. The ELISA allowed the determination of the protein in the concentration range 0.2-25 mu g/l. Measurement of the neutrophil lipocalin concentration showed that human plasma of healthy donors contained 97 +/- 81 mu g/l (n = 122) and that the concentrations in serum were significantly higher (P < 0,001) with 133 +/- 90 mu g/l (n = 122). Neutrophil lipocalin was also found in the urine of healthy donors (8.1 mu g/l; n = 9). Very high concentrations of this lipocalin were found in the synovial fluids of patients suffering from inflammatory rheumatoid arthritis (1.7 +/- 1.4 mg/l; n = 37).