TY - JOUR
T1 - Protein attachment to nanoporous anodic alumina for biotechnological applications
T2 - Influence of pore size, protein size and functionalization path
AU - Baranowska, Malgorzata
AU - Slota, Agata J.
AU - Eravuchira, Pinkie J.
AU - Macias, Gerard
AU - Xifré-Pérez, Elisabet
AU - Pallares, Josep
AU - Ferré-Borrull, Josep
AU - Marsal, Lluís F.
N1 - Funding Information:
This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO) under grant number TEC2012-34397 and by Catalan Government under project AGAUR 2014 SGR 1344 .
PY - 2014/10/1
Y1 - 2014/10/1
N2 - Nanoporous anodic alumina (NAA) is a material with great interest in nanotechnology and with promising applications to biotechnology. Obtaining specific and regularly functionalized NAA surfaces is essential to obtain meaningful results and applications. Silane-PEG-NHS (triethoxysilane-polyethylene-glycol-N-hydroxysuccinimide) is a covalent linker commonly used for single-molecule studies. We investigate the functionalization of NAA with silane-PEG-NHS and compared with two common, but not single-molecule, grafting agents, APTMS (3-aminopropylotrimethoxysilane) as an electrostatic linker, and APTMS-GTA (3-aminopropylotrimethoxysilane-glutaraldehyde) as covalent. Another outcome of this study is to show how two proteins (collagen and bovine serum albumin, BSA) with different properties differentially arrange for different functionalizations and NAA pore sizes. FTIR is used to demonstrate the surface modification steps and fluorescence confocal microscopy reveals that silane-PEG-NHS results in a more homogeneous protein distribution in comparison to the other linkers. Reflection interference Fourier transform spectroscopy confirms the confocal fluorescence microscopy results and permits to estimate the amounts of linker and linked proteins within the pores. These results permit to obtain uniformly chemical modified NAA supports with a great value in biosensing, drug delivery and cell biology.
AB - Nanoporous anodic alumina (NAA) is a material with great interest in nanotechnology and with promising applications to biotechnology. Obtaining specific and regularly functionalized NAA surfaces is essential to obtain meaningful results and applications. Silane-PEG-NHS (triethoxysilane-polyethylene-glycol-N-hydroxysuccinimide) is a covalent linker commonly used for single-molecule studies. We investigate the functionalization of NAA with silane-PEG-NHS and compared with two common, but not single-molecule, grafting agents, APTMS (3-aminopropylotrimethoxysilane) as an electrostatic linker, and APTMS-GTA (3-aminopropylotrimethoxysilane-glutaraldehyde) as covalent. Another outcome of this study is to show how two proteins (collagen and bovine serum albumin, BSA) with different properties differentially arrange for different functionalizations and NAA pore sizes. FTIR is used to demonstrate the surface modification steps and fluorescence confocal microscopy reveals that silane-PEG-NHS results in a more homogeneous protein distribution in comparison to the other linkers. Reflection interference Fourier transform spectroscopy confirms the confocal fluorescence microscopy results and permits to estimate the amounts of linker and linked proteins within the pores. These results permit to obtain uniformly chemical modified NAA supports with a great value in biosensing, drug delivery and cell biology.
KW - BSA
KW - Collagen
KW - GTA
KW - Nanoporous anodic alumina
KW - Reflection interference fourier transform spectroscopy (RIFTS)
KW - Silane-PEG-NHS
UR - http://www.scopus.com/inward/record.url?scp=84905378339&partnerID=8YFLogxK
U2 - 10.1016/j.colsurfb.2014.07.027
DO - 10.1016/j.colsurfb.2014.07.027
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C2 - 25086305
AN - SCOPUS:84905378339
SN - 0927-7765
VL - 122
SP - 375
EP - 383
JO - Colloids and Surfaces B: Biointerfaces
JF - Colloids and Surfaces B: Biointerfaces
ER -