دورية أكاديمية
Liposome Artificial Membrane Permeability Assay by MALDI-hydrogen-deuterium exchange mass spectrometry for peptides and small proteins.
| العنوان: | Liposome Artificial Membrane Permeability Assay by MALDI-hydrogen-deuterium exchange mass spectrometry for peptides and small proteins. |
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| المؤلفون: | Makarov AA; Merck & Co., Inc., MRL, Analytical Research & Development /Process Research & Development, 126 E. Lincoln Ave., Rahway, NJ, 07065, USA. Electronic address: alexey.makarov@merck.com., Pirrone GF; Merck & Co., Inc., MRL, Analytical Research & Development /Process Research & Development, 126 E. Lincoln Ave., Rahway, NJ, 07065, USA., Shchurik V; Merck & Co., Inc., MRL, Analytical Research & Development /Process Research & Development, 126 E. Lincoln Ave., Rahway, NJ, 07065, USA., Regalado EL; Merck & Co., Inc., MRL, Analytical Research & Development /Process Research & Development, 126 E. Lincoln Ave., Rahway, NJ, 07065, USA., Mangion I; Merck & Co., Inc., MRL, Analytical Research & Development /Process Research & Development, 126 E. Lincoln Ave., Rahway, NJ, 07065, USA. |
| المصدر: | Analytica chimica acta [Anal Chim Acta] 2020 Feb 22; Vol. 1099, pp. 111-118. Date of Electronic Publication: 2019 Sep 25. |
| 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: | Alamethicin/*analysis , Aprotinin/*analysis , Gramicidin/*analysis , Melitten/*analysis , Ubiquitin/*analysis, Hydrogen Deuterium Exchange-Mass Spectrometry ; Liposomes/chemistry ; Membranes, Artificial ; Permeability |
| مستخلص: | The pharmaceutical industry's focus has expanded to include peptide and protein-based therapeutics; however, some analytical challenges have arisen along the way, including the urgent need for fast and robust measurement of the membrane permeability of peptides and small proteins. In this study, a simple and efficient approach that utilizes MALDI-TOF-MS to study peptide and protein permeability through an artificial liposome membrane in conjunction with a differential hydrogen-deuterium exchange (HDX) methodology is described. A non-aqueous (aprotic) matrix was evaluated for use with MALDI sample preparation in order to eliminate undesirable hydrogen-deuterium back-exchange. Peptides and proteins were incubated with liposomes and their penetration into the liposome membrane over time was measured by MALDI-MS. A differential HDX approach was used to distinguish the peptides outside of the liposome from those inside. In this regard, the peptides on the outside of the liposomes were labeled using short exposure to deuterium oxide, while the peptides inside of the liposomes were protected from labeling. Subsequently, the unlabeled versus labeled peak area ratios for peptide and protein samples were compared using MALDI-TOF-MS. In this proof-of-concept study, we developed the Liposome Artificial Membrane Permeability Assay (LAMPA) workflow to study three well-known membrane-active model peptides (melittin, alamethicin, and gramicidin) and two model proteins (aprotinin and ubiquitin). The permeability results obtained from this were corroborated by previously reported data for studied peptides and proteins. The proposed LAMPA by MALDI-HDX-MS can be applied in an ultra-high-throughput manner for studying and rank-ordering membrane permeability of peptides and small proteins. Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2019 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Alamethicin; Aprotinin; Gramicidin; Hydrogen-deuterium exchange; Matrix-assisted laser desorption ionization; Melittin |
| المشرفين على المادة: | 0 (Liposomes) 0 (Membranes, Artificial) 0 (Ubiquitin) 1405-97-6 (Gramicidin) 20449-79-0 (Melitten) 27061-78-5 (Alamethicin) 9087-70-1 (Aprotinin) |
| تواريخ الأحداث: | Date Created: 20200128 Date Completed: 20201112 Latest Revision: 20201112 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1016/j.aca.2019.09.063 |
| PMID: | 31986267 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-4324 |
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| DOI: | 10.1016/j.aca.2019.09.063 |