Organoid Sample Preparation and Extraction for LC-MS Peptidomics

Emily L Miedzybrodzka (1) , Rachel E Foreman (1) , Sam G Galvin (1) , Pierre Larraufie (1) , Amy L George (1) , Deborah A Goldspink (1) , Frank Reimann (1) , Fiona M Gribble (1) , Richard G Kay (1)

1 Institute of Metabolic Science-Metabolic Research Laboratories

This protocol describes the peptidomic analysis of organoid lysates, FACS-purified cell populations, and 2D culture secretions by liquid chromatography mass spectrometry (LC-MS). Currently, most peptides are quantified by ELISA, limiting the peptides that can be studied. However, an LC-MS-based approach allows more peptides to be monitored. Our group has previously used LC-MS for tissue peptidomics and secretion of enteroendocrine peptides from primary culture. Now, we extend the use to organoid models.

Domaines: Sciences du Vivant [q-bio]; Biologie cellulaire

Emily L Miedzybrodzka, Rachel E Foreman, Sam G Galvin, Pierre Larraufie, Amy L George, et al.. Organoid Sample Preparation and Extraction for LC-MS Peptidomics. STAR Protocols, 2020, 1, ⟨10.1016/j.xpro.2020.100164⟩. ⟨hal-03165408⟩