In vitro and in cellulo ApoE particle formation, isolation, and characterization

Karina Lindner; Larissa C van Ek; Khader Awwad; Franziska Hanke; Alla V Korepanova; Vladimir Rybin; Katharina Beckenbauer; Anne-Claude Gavin

doi:10.1016/j.xpro.2022.101894

In vitro and in cellulo ApoE particle formation, isolation, and characterization

STAR Protoc. 2022 Dec 16;3(4):101894. doi: 10.1016/j.xpro.2022.101894. Epub 2022 Dec 2.

Authors

Karina Lindner¹, Larissa C van Ek¹, Khader Awwad², Franziska Hanke², Alla V Korepanova³, Vladimir Rybin⁴, Katharina Beckenbauer⁵, Anne-Claude Gavin⁶

Affiliations

¹ Department of Cell Physiology and Metabolism, Faculty of Medicine, University of Geneva, Rue Michel-Servet 1, 1211 Geneva, Switzerland.
² AbbVie Deutschland GmbH & Co. KG Drug Metabolism and Pharmacokinetics, Knollstrasse, 67061 Ludwigshafen, Germany.
³ AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, USA.
⁴ European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
⁵ AbbVie Deutschland GmbH & Co. KG Neuroscience Discovery, Knollstrasse, 67061 Ludwigshafen, Germany. Electronic address: [email protected].
⁶ Department of Cell Physiology and Metabolism, Faculty of Medicine, University of Geneva, Rue Michel-Servet 1, 1211 Geneva, Switzerland. Electronic address: [email protected].

Abstract

Apolipoprotein E (ApoE) particles are responsible for packing and transporting lipids throughout aqueous environments. We detail steps to assess in vitro particles forming from artificial membranes using right-angle light scattering and to measure their size using dynamic light scattering. We further describe how to generate in cellulo ApoE particles containing triacylglycerol under fatty-acid-induced stress. We also detail steps to isolate them from cell secretome by immunoprecipitation and analyze their lipid cargo by thin-layer chromatography. For complete details on the use and execution of this protocol, please refer to Lindner et al. (2022).¹.

Keywords: Cell Biology; Metabolism; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apolipoproteins E* / chemistry
Fatty Acids*

Substances

Fatty Acids
Apolipoproteins E