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A new enzyme-linked immunosorbent assay system for human serum hepatic triglyceride lipase

机译:一种用于人血清肝甘油三酯脂肪酶的新酶联免疫吸附测定系统

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摘要

There is no established method for measuring human hepatic triglyceride (TG) lipase (HTGL) concentration in serum. In this study, we developed new monoclonal Abs (MoAbs) (9A1 mouse MoAb and 141A1 rat MoAb) that react with HTGL both in serum and in postheparin plasma (PHP) and established a novel ELISA system for measuring serum HTGL and PHP-HTGL concentrations. To confirm the specificity of MoAbs, we performed immunoprecipitation-immunoblotting analysis. Both 9A1 mouse MoAb and 141A1 rat MoAb were able to immunoprecipitate not only recombinant HTGL and PHP-HTGL but also serum HTGL, demonstrating that HTGL exists in serum obtained without heparin injection. This method yielded intra-and interassay coefficients of variation of <6% and showed no cross-reactivity with LPL or endothelial lipase. In clinical analysis on 42 male subjects with coronary artery disease, there were strong positive correlations of serum HTGL concentration to PHPHTGL concentration (r = 0.727, P < 0.01). Serum HTGL concentrations showed positive correlations to serum TGs (r = 0.314, P < 0.05) and alanine aminotransferase (r = 0.406, P < 0.01), and tendencies toward positive correlations to LDL cholesterol, small dense LDL, and gamma GTP.jlr These results suggest that this new ELISA method for measuring serum HTGL is applicable in daily clinical practice.
机译:在血清中没有建立的测量人肝甘油三酯(TG)脂肪酶(HTGL)浓度的方法。在这项研究中,我们开发了新的单克隆ABS(MoAb)(MoAb)(9A1小鼠MoAb和141A1大鼠莫阿布),其在血清中和培磷脂血浆(PHP)中与HTGL反应,并建立了一种用于测量血清HTGL和PHP-HTGL浓度的新型ELISA系统。为了确认莫阿布的特异性,我们进行了免疫沉淀 - 免疫印迹分析。均可不仅可以重组HTGL和PHP-HTGL,而且还能够进行免疫沉淀,但也能够血清HTG1,证明HTGL存在于没有肝素注射的血清中。该方法产生的帧内间变异的系数<6%,并且没有与LPL或内皮脂肪酶的交叉反应性。在42例男性受试者患有冠状动脉疾病的临床分析中,血清HTGL浓度与PHPHTGL浓度有强的正相关(R = 0.727,P <0.01)。血清HTGL浓度显示出与血清TGS(R = 0.314,P <0.05)和丙氨酸氨基转移酶(R = 0.406,P <0.01)的阳性相关性,并趋于与LDL胆固醇,小密集LDL和Gamma GTP.JLR的阳性相关性结果表明,这种新的ELISA测量血清HTGL的方法适用于日常临床实践。

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