دورية أكاديمية
Rapid label-free cell-based Approach Membrane Permeability Assay using MALDI-hydrogen-deuterium exchange mass spectrometry for peptides.
| العنوان: | Rapid label-free cell-based Approach Membrane Permeability Assay using MALDI-hydrogen-deuterium exchange mass spectrometry for peptides. |
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| المؤلفون: | Makarov AA; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA. Electronic address: alexey.makarov@merck.com., Jiang Y; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Sondey C; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Zhang M; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Mansueto MS; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Pirrone GF; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Huang C; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Biswas K; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Duggal R; Merck & Co. Inc., MRL, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA., Al-Sayah MA; Merck & Co. Inc., MRL, 213 E. Grand Avenue, So. San Francisco, CA, 94080, USA., Regalado EL; Merck & Co. Inc., MRL, 126 E. Lincoln Avenue, Rahway, NJ, 07065, USA., Mangion I; Merck & Co. Inc., MRL, 126 E. Lincoln Avenue, Rahway, NJ, 07065, USA. |
| المصدر: | Analytica chimica acta [Anal Chim Acta] 2022 Sep 08; Vol. 1225, pp. 340234. Date of Electronic Publication: 2022 Aug 06. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0370534 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4324 (Electronic) Linking ISSN: 00032670 NLM ISO Abbreviation: Anal Chim Acta Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Original Publication: Amsterdam. |
| مواضيع طبية MeSH: | Deuterium Exchange Measurement*/methods , Hydrogen Deuterium Exchange-Mass Spectrometry*, Cell Membrane Permeability ; Deuterium/chemistry ; Hydrogen/chemistry ; Peptides ; Permeability ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods |
| مستخلص: | Peptide therapeutics are a growing modality in the pharmaceutical industry and expanding these therapeutics to hit intracellular targets would require establishing cell permeability. Rapid measurement target-agnostic cell permeability of peptides is still analytically challenging. In this study, we demonstrate the development of a rapid high-throughput label-free methodology based on a MALDI-hydrogen-deuterium exchange mass spectrometry (MALDI-HDX-MS) approach to rank-order peptide cell membrane permeability using live THP-1 and AsPc-1 cells. Peptides were incubated in the presence of live cells and their permeability into the cells over time was measured by MALDI-HDX-MS. A differential hydrogen-deuterium exchange approach was used to distinguish the peptides outside of the cells from those inside. The peptides on the outside of the cells were labeled using sufficiently short exposure to deuterium oxide, while the peptides inside of the cells were protected from labeling as a result of permeation into the cells. The deuterium labeled and peak area ratios of unlabeled peptides were compared and plotted over time. The developed methodology, referred to as Cell-based Approach Membrane Permeability Assay (CAMPA), was applied to study an array of 24 diverse peptides including cell-penetrating peptides, stapled and macrocyclic peptides. The cell membrane permeability results observed by CAMPA were corroborated by previously reported in literature data. The CAMPA MALDI-MS analysis was fully automated including MS data processing using internally developed Python scripts. Moreover, CAMPA was demonstrated to be useful for differentiating passive and active cell transportation by using an endocytosis inhibitor in cell incubation media for selected peptides. (Copyright © 2022 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: AsPc-1; CAMPA; HDX; MALDI; THP-1 |
| المشرفين على المادة: | 0 (Peptides) 7YNJ3PO35Z (Hydrogen) AR09D82C7G (Deuterium) |
| تواريخ الأحداث: | Date Created: 20220829 Date Completed: 20220831 Latest Revision: 20220831 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/j.aca.2022.340234 |
| PMID: | 36038238 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-4324 |
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| DOI: | 10.1016/j.aca.2022.340234 |