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

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...

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Autores principales: Kluge, Stefan, Boermel, Lisa, Schubert, Martin, Lorkowski, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7132154/
https://www.ncbi.nlm.nih.gov/pubmed/32274337
http://dx.doi.org/10.1016/j.mex.2020.100865
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author Kluge, Stefan
Boermel, Lisa
Schubert, Martin
Lorkowski, Stefan
author_facet Kluge, Stefan
Boermel, Lisa
Schubert, Martin
Lorkowski, Stefan
author_sort Kluge, Stefan
collection PubMed
description 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 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.
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spelling pubmed-71321542020-04-09 Simple and rapid real-time monitoring of LPL activity in vitro Kluge, Stefan Boermel, Lisa Schubert, Martin Lorkowski, Stefan MethodsX Pharmacology, Toxicology and Pharmaceutical Science 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 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. Elsevier 2020-03-17 /pmc/articles/PMC7132154/ /pubmed/32274337 http://dx.doi.org/10.1016/j.mex.2020.100865 Text en © 2020 The Authors. Published by Elsevier B.V. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Pharmacology, Toxicology and Pharmaceutical Science
Kluge, Stefan
Boermel, Lisa
Schubert, Martin
Lorkowski, Stefan
Simple and rapid real-time monitoring of LPL activity in vitro
title Simple and rapid real-time monitoring of LPL activity in vitro
title_full Simple and rapid real-time monitoring of LPL activity in vitro
title_fullStr Simple and rapid real-time monitoring of LPL activity in vitro
title_full_unstemmed Simple and rapid real-time monitoring of LPL activity in vitro
title_short Simple and rapid real-time monitoring of LPL activity in vitro
title_sort simple and rapid real-time monitoring of lpl activity in vitro
topic Pharmacology, Toxicology and Pharmaceutical Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7132154/
https://www.ncbi.nlm.nih.gov/pubmed/32274337
http://dx.doi.org/10.1016/j.mex.2020.100865
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