Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries

The discovery of new membrane-active peptides (MAPs) is an area of considerable interest in modern biotechnology considering their ample applicability in several fields ranging from the development of novel delivery vehicles (via cell-penetrating peptides) to responding to the latent threat of antib...

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Autores principales:	Saúl C. Gómez, Valentina Quezada, Isabella Quiroz, Carolina Muñoz-Camargo, Johann F. Osma, Luis H. Reyes, Juan C. Cruz
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Giant Unilamellar Vesicles micromixers multiphysics simulation chitosan microparticles Mechanical engineering and machinery TJ1-1570
Acceso en línea:	https://doaj.org/article/f127e641b2654e889a0f7551f9eafc1e
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spelling	oai:doaj.org-article:f127e641b2654e889a0f7551f9eafc1e2021-11-25T18:23:31ZDesign and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries10.3390/mi121113772072-666Xhttps://doaj.org/article/f127e641b2654e889a0f7551f9eafc1e2021-11-01T00:00:00Zhttps://www.mdpi.com/2072-666X/12/11/1377https://doaj.org/toc/2072-666XThe discovery of new membrane-active peptides (MAPs) is an area of considerable interest in modern biotechnology considering their ample applicability in several fields ranging from the development of novel delivery vehicles (via cell-penetrating peptides) to responding to the latent threat of antibiotic resistance (via antimicrobial peptides). Different strategies have been devised for such discovery process, however, most of them involve costly, tedious, and low-efficiency methods. We have recently proposed an alternative route based on constructing a non-rationally designed library recombinantly expressed on the yeasts’ surfaces. However, a major challenge is to conduct a robust and high-throughput screening of possible candidates with membrane activity. Here, we addressed this issue by putting forward low-cost microfluidic platforms for both the synthesis of Giant Unilamellar Vesicles (GUVs) as mimicking entities of cell membranes and for providing intimate contact between GUVs and homologues of yeasts expressing MAPs. The homologues were chitosan microparticles functionalized with the membrane translocating peptide Buforin II, while intimate contact was through passive micromixers with different channel geometries. Both microfluidic platforms were evaluated both in silico (via Multiphysics simulations) and in vitro with a high agreement between the two approaches. Large and stable GUVs (5–100 µm) were synthesized effectively, and the mixing processes were comprehensively studied leading to finding the best operating parameters. A serpentine micromixer equipped with circular features showed the highest average encapsulation efficiencies, which was explained by the unique mixing patterns achieved within the device. The microfluidic devices developed here demonstrate high potential as platforms for the discovery of novel MAPs as well as for other applications in the biomedical field such as the encapsulation and controlled delivery of bioactive compounds.Saúl C. GómezValentina QuezadaIsabella QuirozCarolina Muñoz-CamargoJohann F. OsmaLuis H. ReyesJuan C. CruzMDPI AGarticleGiant Unilamellar Vesiclesmicromixersmultiphysics simulationchitosan microparticlesMechanical engineering and machineryTJ1-1570ENMicromachines, Vol 12, Iss 1377, p 1377 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Giant Unilamellar Vesicles micromixers multiphysics simulation chitosan microparticles Mechanical engineering and machinery TJ1-1570
spellingShingle	Giant Unilamellar Vesicles micromixers multiphysics simulation chitosan microparticles Mechanical engineering and machinery TJ1-1570 Saúl C. Gómez Valentina Quezada Isabella Quiroz Carolina Muñoz-Camargo Johann F. Osma Luis H. Reyes Juan C. Cruz Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
description	The discovery of new membrane-active peptides (MAPs) is an area of considerable interest in modern biotechnology considering their ample applicability in several fields ranging from the development of novel delivery vehicles (via cell-penetrating peptides) to responding to the latent threat of antibiotic resistance (via antimicrobial peptides). Different strategies have been devised for such discovery process, however, most of them involve costly, tedious, and low-efficiency methods. We have recently proposed an alternative route based on constructing a non-rationally designed library recombinantly expressed on the yeasts’ surfaces. However, a major challenge is to conduct a robust and high-throughput screening of possible candidates with membrane activity. Here, we addressed this issue by putting forward low-cost microfluidic platforms for both the synthesis of Giant Unilamellar Vesicles (GUVs) as mimicking entities of cell membranes and for providing intimate contact between GUVs and homologues of yeasts expressing MAPs. The homologues were chitosan microparticles functionalized with the membrane translocating peptide Buforin II, while intimate contact was through passive micromixers with different channel geometries. Both microfluidic platforms were evaluated both in silico (via Multiphysics simulations) and in vitro with a high agreement between the two approaches. Large and stable GUVs (5–100 µm) were synthesized effectively, and the mixing processes were comprehensively studied leading to finding the best operating parameters. A serpentine micromixer equipped with circular features showed the highest average encapsulation efficiencies, which was explained by the unique mixing patterns achieved within the device. The microfluidic devices developed here demonstrate high potential as platforms for the discovery of novel MAPs as well as for other applications in the biomedical field such as the encapsulation and controlled delivery of bioactive compounds.
format	article
author	Saúl C. Gómez Valentina Quezada Isabella Quiroz Carolina Muñoz-Camargo Johann F. Osma Luis H. Reyes Juan C. Cruz
author_facet	Saúl C. Gómez Valentina Quezada Isabella Quiroz Carolina Muñoz-Camargo Johann F. Osma Luis H. Reyes Juan C. Cruz
author_sort	Saúl C. Gómez
title	Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
title_short	Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
title_full	Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
title_fullStr	Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
title_full_unstemmed	Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries
title_sort	design and manufacture of a low-cost microfluidic system for the synthesis of giant liposomes for the encapsulation of yeast homologues: applications in the screening of membrane-active peptide libraries
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f127e641b2654e889a0f7551f9eafc1e
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Design and Manufacture of a Low-Cost Microfluidic System for the Synthesis of Giant Liposomes for the Encapsulation of Yeast Homologues: Applications in the Screening of Membrane-Active Peptide Libraries

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