Kluge Stefan, Boermel Lisa, Schubert Martin, Lorkowski Stefan
Institute of Nutritional Sciences, Friedrich Schiller University Jena, Germany.
Competence Cluster for Nutrition and Cardiovascular Health (nutriCARD) Halle-Jena-Leipzig, Germany.
MethodsX. 2020 Mar 17;7:100865. doi: 10.1016/j.mex.2020.100865. eCollection 2020.
Since elevated plasma triglycerides are an independent risk factor for cardiovascular diseases, lipoprotein lipase (LPL) is an interesting target for drug development. However, investigation of LPL remains challenging, as most of the commercially available assays are limited to the determination of LPL activity. Thus, we focused on the evaluation of a simple real-time fluorescence assay for the measurement of LPL activity that can be combined with additional cell or molecular biological assays in the same cell sample. Our procedure allows for a more comprehensive characterization of potential regulatory compounds targeting the LPL system. The presented assay procedure provides several advantages over currently available commercial LPL activity assays:1.12-well cell culture plate design for the simultaneous investigation of up to three different compounds of interest (including all assay controls).2.24 h real-time acquisition of LPL activity for the identification of the optimal time point for further measurements.3.Measurement of LPL activity can be supplemented by additional cell or molecular biological assays in the same cell sample.
由于血浆甘油三酯升高是心血管疾病的独立危险因素，脂蛋白脂肪酶（LPL）是药物开发的一个有趣靶点。然而，对LPL的研究仍然具有挑战性，因为大多数市售检测方法仅限于测定LPL活性。因此，我们专注于评估一种简单的实时荧光检测方法来测量LPL活性，该方法可与同一细胞样本中的其他细胞或分子生物学检测方法相结合。我们的方法能够更全面地表征靶向LPL系统的潜在调节化合物。与目前可用的商业LPL活性检测方法相比，本检测方法具有以下几个优点：1.采用12孔细胞培养板设计，可同时研究多达三种不同的目标化合物（包括所有检测对照）。2.对LPL活性进行24小时实时采集，以确定进一步测量的最佳时间点。3.在同一细胞样本中，LPL活性的测量可通过额外的细胞或分子生物学检测进行补充。
