Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/103225
Title: Chemically synthesized peptide libraries as a new source of BBB shuttles. Use of mass spectrometry for peptide identification
Author: Guixer, Bernat
Arroyo, X.
Belda, I.
Sabidó Aguadé, Eduard
Teixidó Turà, Meritxell
Giralt Lledó, Ernest
Keywords: Síntesi de pèptids
Espectrometria de masses
Barrera hematoencefàlica
Peptide synthesis
Mass spectrometry
Blood-brain barrier
Issue Date: 20-Jul-2016
Publisher: John Wiley & Sons
Abstract: The blood-brain barrier (BBB) is a biological barrier that protects the brain from neurotoxic agents and regulates the influx and efflux of molecules required for its correct function. This stringent regulation hampers the passage of brain parenchyma-targeting drugs across the BBB. BBB shuttles have been proposed as a way to overcome this hurdle because these peptides can not only cross the BBB but also carry molecules which would otherwise be unable to cross the barrier unaided. Here we developed a new high-throughput screening methodology to identify new peptide BBB shuttles in a broadly unexplored chemical space. By introducing d-amino acids, this approach screens only protease-resistant peptides. This methodology combines combinatorial chemistry for peptide library synthesis, in vitro models mimicking the BBB for library evaluation and state-of-the-art mass spectrometry techniques to identify those peptides able to cross the in vitro assays. BBB shuttle synthesis was performed by the mix-and-split technique to generate a library based on the following: Ac-d-Arg-XXXXX-NH2, where X were: d-Ala (a), d-Arg (r), d-Ile (i), d-Glu (e), d-Ser (s), d-Trp (w) or d-Pro (p). The assays used comprised the in vitro cell-based BBB assay (mimicking both active and passive transport) and the PAMPA (mimicking only passive diffusion). The identification of candidates was determined using a two-step mass spectrometry approach combining LTQ-Orbitrap and Q-trap mass spectrometers. Identified sequences were postulated to cross the BBB models. We hypothesized that some sequences cross the BBB through passive diffusion mechanisms and others through other mechanisms, including paracellular flux and active transport. These results provide a new set of BBB shuttle peptide families. Furthermore, the methodology described is proposed as a consistent approach to search for protease-resistant therapeutic peptides
Note: Versió postprint del document publicat a: https://doi.org/10.1002/psc.2900
It is part of: Journal of Peptide Science, 2016, vol. 22, p. 577-591
Related resource: https://doi.org/10.1002/psc.2900
URI: http://hdl.handle.net/2445/103225
ISSN: 1075-2617
Appears in Collections:Articles publicats en revistes (Química Inorgànica i Orgànica)
Articles publicats en revistes (Institut de Recerca Biomèdica (IRB Barcelona))

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