Microfluidic biofunctionalisation protocols to form multi-valent interactions for cell rolling and phenotype modification investigations

Handle URI:
http://hdl.handle.net/10754/562840
Title:
Microfluidic biofunctionalisation protocols to form multi-valent interactions for cell rolling and phenotype modification investigations
Authors:
Perozziello, Gerardo; Simone, Giuseppina; Malara, Natalia Maria; La Rocca, Rosanna; Tallerico, Rossana; Catalano, Rossella; Pardeo, Francesca; Candeloro, Patrizio; Cuda, Giovanni; Carbone, Ennio; Di Fabrizio, Enzo M. ( 0000-0001-5886-4678 )
Abstract:
In this study, we propose a fast, simple method to biofunctionalise microfluidic systems for cellomic investigations based on micro-fluidic protocols. Many available processes either require expensive and time-consuming protocols or are incompatible with the fabrication of microfluidic systems. Our method differs from the existing since it is applicable to an assembled system, uses few microlitres of reagents and it is based on the use of microbeads. The microbeads have specific surface moieties to link the biomolecules and couple cell receptors. Furthermore, the microbeads serve as arm spacer and offer the benefit of the multi-valent interaction. Microfluidics was adapted together with topology and biochemistry surface modifications to offer the microenvironment for cellomic studies. Based on this principle, we exploit the streptavidin-biotin interaction to couple antibodies to the biofunctionalised microfluidic environment within 5 h using 200 μL of reagents and biomolecules. We selected the antibodies able to form complexes with the MHC class I (MHC-I) molecules present on the cell membrane and involved in the immune surveillance. To test the microfluidic system, tumour cell lines (RMA) were rolled across the coupled antibodies to recognise and strip MHC-I molecules. As result, we show that cell rolling performed inside a microfluidic chamber functionalised with beads and the opportune antibody facilitate the removal of MHC class I molecules. We showed that the level of median fluorescent intensity of the MHC-I molecules is 300 for cells treated in a not biofunctionalised surface. It decreased to 275 for cells treated in a flat biofunctionalised surface and to 250 for cells treated on a surface where biofunctionalised microbeads were immobilised. The cells with reduced expression of MHC-I molecules showed, after cytotoxicity tests, susceptibility 3.5 times higher than normal cells. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Physical Sciences and Engineering (PSE) Division; Materials Science and Engineering Program
Publisher:
Wiley-Blackwell
Journal:
ELECTROPHORESIS
Issue Date:
Jul-2013
DOI:
10.1002/elps.201300106
PubMed ID:
23616364
Type:
Article
ISSN:
01730835
Sponsors:
This work was partially supported by the project PON "Nuove strategie nanotecnologiche per la messa a punto di farmaci e presidi diagnostici diretti verso cellule cancerose circolanti" (code: PON01_02782) and the project FIRB "Rete Nazionale di Ricerca sulle Nanoscienze ItalNanoNet" (cod. RBPR05JH2P_010, CUP B41J09000110005) granted to the nanotechnology laboratory of the Department of Experimental Medicine of the University of Magna Graecia of Catanzaro. Ennio Carbone is Wenner Gren Foundation senior research fellow and was supported by grant from Italian Association for Cancer Research (AIRC IG 10189). Rossana Tallerico is awarded by triennial fellowships "Luciana Selce" FIRC. The authors have declared no conflict of interest.
Appears in Collections:
Articles; Physical Sciences and Engineering (PSE) Division; Materials Science and Engineering Program; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorPerozziello, Gerardoen
dc.contributor.authorSimone, Giuseppinaen
dc.contributor.authorMalara, Natalia Mariaen
dc.contributor.authorLa Rocca, Rosannaen
dc.contributor.authorTallerico, Rossanaen
dc.contributor.authorCatalano, Rossellaen
dc.contributor.authorPardeo, Francescaen
dc.contributor.authorCandeloro, Patrizioen
dc.contributor.authorCuda, Giovannien
dc.contributor.authorCarbone, Ennioen
dc.contributor.authorDi Fabrizio, Enzo M.en
dc.date.accessioned2015-08-03T11:12:16Zen
dc.date.available2015-08-03T11:12:16Zen
dc.date.issued2013-07en
dc.identifier.issn01730835en
dc.identifier.pmid23616364en
dc.identifier.doi10.1002/elps.201300106en
dc.identifier.urihttp://hdl.handle.net/10754/562840en
dc.description.abstractIn this study, we propose a fast, simple method to biofunctionalise microfluidic systems for cellomic investigations based on micro-fluidic protocols. Many available processes either require expensive and time-consuming protocols or are incompatible with the fabrication of microfluidic systems. Our method differs from the existing since it is applicable to an assembled system, uses few microlitres of reagents and it is based on the use of microbeads. The microbeads have specific surface moieties to link the biomolecules and couple cell receptors. Furthermore, the microbeads serve as arm spacer and offer the benefit of the multi-valent interaction. Microfluidics was adapted together with topology and biochemistry surface modifications to offer the microenvironment for cellomic studies. Based on this principle, we exploit the streptavidin-biotin interaction to couple antibodies to the biofunctionalised microfluidic environment within 5 h using 200 μL of reagents and biomolecules. We selected the antibodies able to form complexes with the MHC class I (MHC-I) molecules present on the cell membrane and involved in the immune surveillance. To test the microfluidic system, tumour cell lines (RMA) were rolled across the coupled antibodies to recognise and strip MHC-I molecules. As result, we show that cell rolling performed inside a microfluidic chamber functionalised with beads and the opportune antibody facilitate the removal of MHC class I molecules. We showed that the level of median fluorescent intensity of the MHC-I molecules is 300 for cells treated in a not biofunctionalised surface. It decreased to 275 for cells treated in a flat biofunctionalised surface and to 250 for cells treated on a surface where biofunctionalised microbeads were immobilised. The cells with reduced expression of MHC-I molecules showed, after cytotoxicity tests, susceptibility 3.5 times higher than normal cells. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.en
dc.description.sponsorshipThis work was partially supported by the project PON "Nuove strategie nanotecnologiche per la messa a punto di farmaci e presidi diagnostici diretti verso cellule cancerose circolanti" (code: PON01_02782) and the project FIRB "Rete Nazionale di Ricerca sulle Nanoscienze ItalNanoNet" (cod. RBPR05JH2P_010, CUP B41J09000110005) granted to the nanotechnology laboratory of the Department of Experimental Medicine of the University of Magna Graecia of Catanzaro. Ennio Carbone is Wenner Gren Foundation senior research fellow and was supported by grant from Italian Association for Cancer Research (AIRC IG 10189). Rossana Tallerico is awarded by triennial fellowships "Luciana Selce" FIRC. The authors have declared no conflict of interest.en
dc.publisherWiley-Blackwellen
dc.subjectBiofunctionalisationen
dc.subjectCell rollingen
dc.subjectMulti-valent interactionsen
dc.subjectPhenotype modificationsen
dc.titleMicrofluidic biofunctionalisation protocols to form multi-valent interactions for cell rolling and phenotype modification investigationsen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentPhysical Sciences and Engineering (PSE) Divisionen
dc.contributor.departmentMaterials Science and Engineering Programen
dc.identifier.journalELECTROPHORESISen
dc.contributor.institutionDepartment of Experimental Medicine, Bio Nano Engineering and Technology for Medicine (BioNEM) Laboratory, University Magna Graecia of Catanzaro, Catanzaro, Italyen
dc.contributor.institutionInterdisciplinary Research Centre in Biomaterials, CRIB-University of Napoli Federico II, Napoli, Italyen
dc.contributor.institutionNanostructures Group, Italian Institute of Technology IIT, Genova, Italyen
dc.contributor.institutionDepartment of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Swedenen
kaust.authorDi Fabrizio, Enzo M.en

Related articles on PubMed

All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.