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用于测定脂蛋白脂肪酶质量浓度的ELISA方法的建立与评估——与一种商业化的一步酶免疫测定法的比较

Development and evaluation of an ELISA method for the determination of lipoprotein lipase mass concentration--comparison with a commercial, one-step enzyme immunoassay.

作者信息

Antikainen M, Suurinkeroinen L, Jauhiainen M, Ehnholm C, Taskinen M R

机构信息

Children Hospital, University of Helsinki, Finland.

出版信息

Eur J Clin Chem Clin Biochem. 1996 Jul;34(7):547-53. doi: 10.1515/cclm.1996.34.7.547.

Abstract

We developed a non-competitive, enzyme-linked, immunosorbent assay (ELISA) for the quantitation of lipoprotein lipase (LPL) in human postheparin plasma using affinity-purified antihuman milk lipoprotein lipase antibodies produced in chicken eggs and a monoclonal antibody directed against human lipoprotein lipase. We compared our ELISA method with a commercially available sandwich-enzyme immunoassay (Markit-F LPL EIA Kit, Dainippon Pharmaceutical Co, Ltd. Osaka, Japan). The reference values for lipoprotein lipase catalytic activity concentration and mass concentration in healthy Finns were determined. Lipoprotein lipase activity concentration (mean +/- SD) was 297 +/- 112 U/l in women, and mass concentration as measured by the ELISA method was 1058 +/- 367 micrograms/l. The corresponding values for men were 247 +/- 97 U/l and 815 +/- 207 micrograms/l, respectively. Across the whole concentration range of the ELISA method, the control samples' intra- and inter-assay coefficients of variation (CV) were 5.1% and 6.5%, respectively. The correlation between the ELISA and EIA methods was good, r = +0.81. The importance of the correct standardisation of immunoassays is discussed.

摘要

我们开发了一种非竞争性酶联免疫吸附测定法(ELISA),用于定量人肝素后血浆中的脂蛋白脂肪酶(LPL),该方法使用了用鸡蛋制备的亲和纯化抗人乳脂蛋白脂肪酶抗体和一种针对人脂蛋白脂肪酶的单克隆抗体。我们将我们的ELISA方法与一种市售的夹心酶免疫测定法(Markit-F LPL EIA试剂盒,日本大阪大日本制药株式会社)进行了比较。测定了健康芬兰人脂蛋白脂肪酶催化活性浓度和质量浓度的参考值。女性脂蛋白脂肪酶活性浓度(平均值±标准差)为297±112 U/l,ELISA法测得的质量浓度为1058±367μg/l。男性的相应值分别为247±97 U/l和815±207μg/l。在ELISA方法的整个浓度范围内,对照样品的批内和批间变异系数(CV)分别为5.1%和6.5%。ELISA法和EIA法之间的相关性良好,r = +0.81。讨论了免疫测定正确标准化的重要性。

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