title: Electrospray deposition in vacuum as method to create functionally active protein immobilization on polymeric substrates.
abstract: We demonstrate in this work the deposition of a large biological molecule (fibronectin) on polymeric substrates in a high vacuum environment using an electrospray deposition system. Fibronectin was deposited and its distribution and structure investigated and retention of function (ability to promote cell adhesion) on return to liquid environment is shown. AFM was used to monitor changes in the morphology of the surface before and after fibronectin deposition, whilst the biological activity of the deposited protein is assessed through a quantitative analysis of the biomolecular adhesion and migration of fibroblast cells to the modified surfaces. For the first time we have demonstrated that using high vacuum electrospray deposition it is possible to deposit large protein molecules on polymeric surfaces whilst maintaining the protein activity. The deposition of biological molecules such as proteins with the retention of their activity onto clean well-controlled surfaces under vacuum condition, offers the possibility for future studies utilizing high resolution vacuum based techniques at the atomic and molecular scale providing a greater understanding of protein-surface interface behaviour of relevance to a wide range of applications such as in sensors, diagnostics and tissue engineering.
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, The University of Nottingham, Nottingham NG7 2RD, UK. Electronic address: enzofornari@hotmail.co.uk.
authors.forename: Enzo
authors.initials: E
authors.lastname: Fornari
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, The University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Clive J
authors.initials: CJ
authors.lastname: Roberts
authors.affiliation: School of Physics and Astronomy and Nottingham Nanotechnology and Nanoscience Centre (NNNC), University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Robert H
authors.initials: RH
authors.lastname: Temperton
authors.affiliation: School of Physics and Astronomy and Nottingham Nanotechnology and Nanoscience Centre (NNNC), University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: James N
authors.initials: JN
authors.lastname: O'Shea
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000378:metabolism; D002448:Cell Adhesion; D002465:Cell Movement; D005347:Fibroblasts / Q000166:cytology; D005353:Fibronectins / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D011108:Polymers / Q000737:chemistry; D013499:Surface Properties; D014618:Vacuum
chemical_list: D001672:Biocompatible Materials; D005353:Fibronectins; D055767:Immobilized Proteins; D011108:Polymers
keywords: Cell adhesion and migration; Fibronectin deposition; Protein activity; Vacuum electrospray deposition