Simple and rapid real-time monitoring of LPL activity in vitro

• Published in: MethodsX 2020-01, Vol.7, p.100865-100865, Article 100865
• Main Authors: Kluge, Stefan, Boermel, Lisa, Schubert, Martin, Lorkowski, Stefan
• Format: Article
• Language: English
• Subjects: Fluorescence; Lipoprotein lipase (LPL); LPL activity assay; Pharmacology, Toxicology and Pharmaceutical Science; Real-time assay
• ISSN: 2215-0161; 2215-0161
• DOI: 10.1016/j.mex.2020.100865

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 in vitro 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 in vitro 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. [Display omitted]