Spectroscopic and microscopic characterization of biosensor surfaces with protein/amino-organosilane/silicon structure

被引：34

作者：

Awsiuk, K. ^{[1
]}

Bernasik, A. ^{[2
]}

Kitsara, M. ^{[3
]}

Budkowski, A. ^{[1
]}

Petrou, P. ^{[3
]}

Kakabakos, S. ^{[3
]}

Prauzner-Bechcicki, S. ^{[1
]}

Rysz, J. ^{[1
]}

Raptis, I. ^{[4
]}

机构：

[1] Jagiellonian Univ, M Smoluchowski Inst Phys, PL-30059 Krakow, Poland

[2] AGH Univ Sci & Technol, Fac Phys & Appl Comp Sci, PL-30059 Krakow, Poland

[3] NCSR Demokritos, Inst Radioisotopes & Radiodiagnost Prod, Aghia Paraskevi 15310, Greece

[4] NCSR Demokritos, Inst Microelect, Aghia Paraskevi 15310, Greece

来源：

COLLOIDS AND SURFACES B-BIOINTERFACES | 2012年 / 90卷

关键词：

Immunoglobulins; Amino-organosilane films; Specific binding; Angle-resolved X-ray photoelectron spectroscopy; Atomic force microscopy; Near-field scanning optical microscopy; ATOMIC-FORCE MICROSCOPY; SELF-ASSEMBLED MONOLAYERS; ADSORPTION; XPS; ORIENTATION; BINDING; QUARTZ; IGG; AFM;

D O I：

10.1016/j.colsurfb.2011.10.017

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Composition and structure of biorecognition protein layers created on silicon substrates modified with amino-organosilanes determine the sensitivity and specificity of silicon based biosensing devices. In the present work, diverse spectroscopic and microscopic methods were applied to characterize model biosensor surfaces, formed on Si3N4 or SiO2 by modification with (3-aminopropyl)triethoxysilane, coating with rabbit gamma-globulins (IgGs) through physical adsorption, blocking with bovine serum albumin (BSA) and specific binding of an anti-rabbit IgG antibody. In addition, silanized substrates with directly adsorbed BSA or anti-rabbit IgG antibody were examined as reference surfaces. The protein/amino-organosilane/silicon structure of all surfaces was confirmed by X-ray photoelectron spectroscopy. Homogeneity of protein coverage was verified with near-field scanning optical microscope, working in reflection and fluorescence mode. Surface coverage with proteins was determined with angle-resolved XPS using a previously established bilayer approach. Inner structure of protein layers was examined with atomic force microscopy. Vertical arrangement of carbon functional groups was revealed by high resolution ARXPS. Combined spectroscopic and microscopic data reveal the complex character of interactions with the immobilized IgG molecules during blocking with BSA and immunoreaction with anti-IgG antibody. Within experimental error, neither surface coverage nor lateral structural scales of protein layer (provided by Fourier and auto-correlation analysis of topographic and phase images) increase during blocking procedure. On the other hand, coverage and all structural measures rise considerably after immunoreaction. In addition, it was found that polar functional groups orient towards substrate for all protein layers, independently of coverage, prior to and after both blocking and specific binding. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：159 / 168

页数：10

共 35 条

[1] Can scanning near-field optical microscopy be compared with confocal laser scanning microscopy?: A preliminary study on α-sarcoglycan and β1D-integrin in human skeletal muscle [J].