Applied Surface Science, 325 (2015) 91-99

Biosensing interfaces consisting of linker molecules (COOH or NH2) and charged, antifouling moieties (( SO3− and N(Me)3) for biosensing applications were prepared for the first time by the in situ deposition of mixtures of aryl diazonium cations on indium tin oxide (ITO) electrodes. A linker molecule is required for the attachment of biorecognition molecules (e.g., antibodies, enzymes, DNA chains, and aptamers) close to the transducer surface. The attached molecules improve the biosensing sensitivity and also provide a short response time for analyte detection. Thus, the incorporation of a linker and antifouling molecules is an important interfacial design for both affinity and enzymatic biosensors. The reductive adsorption behavior and electrochemical measurement were studied for (1) an individual compound and (2) a mixture of antifouling zwitterionic molecules together with linker molecules [combination 1: 4sulfophenyl (SP), 4-trimethylammoniophenyl (TMAP), and 1,4-phenylenediamine (PPD); combination 2: 4-sulfophenyl (SP), 4-trimethylammoniophenyl (TMAP), and 4-aminobenzoic acid (PABA)] of aryl diazonium cations grafted onto an ITO electrode. The mixture ratios of SP:TMAP:PPD and SP:TMAP:PABA that provided the greatest resistance to non-specific protein adsorptions of bovine serum albumin labeled with fluorescein isothiocyanate (BSA–FITC) and cytochrome c labeled with rhodamine B isothiocyanate (RBITC–Cyt c) were determined by confocal laser scanning microscopy (CLSM). For the surface antifouling study, we used 2-[2-(2-methoxyethoxy) ethoxy]acetic acid (OEG) as a standard control because of its prominent antifouling properties. Surface compositions of combinations 1 and 2 were characterized Applied Surface Science 325 (2015) 91–99 Post-print version using X-ray photoelectron spectroscopy (XPS). Field-emission scanning electron microscopy (FE-SEM) was used to characterize the morphology of the grafted films to confirm the even distribution between linker and antifouling molecules grafted onto the ITO surfaces. Combination 1 (SP:TMAP:PPD) with a ratio of 0.5:1.5:0.37 exhibited the best antifouling capability with respect to resisting the nonspecific adsorption of proteins.

[1]  M. Trau,et al.  Femtomolar detection of a cancer biomarker protein in serum with ultralow background current by anodic stripping voltammetry. , 2012, Chemical communications.

[2]  S Zhang,et al.  Materials and techniques for electrochemical biosensor design and construction. , 2000, Biosensors & bioelectronics.

[3]  Jong Il Rhee,et al.  Anti-fouling epoxy coatings for optical biosensor application based on phosphorylcholine , 2007 .

[4]  R. F. Dutra,et al.  A sensor tip based on carbon nanotube-ink printed electrode for the dengue virus NS1 protein. , 2013, Biosensors & bioelectronics.

[5]  George M. Whitesides,et al.  Redox Properties of Cytochrome c Adsorbed on Self-Assembled Monolayers: A Probe for Protein Conformation and Orientation , 2002 .

[6]  Alexander Revzin,et al.  Micropatterning of proteins and mammalian cells on indium tin oxide. , 2009, ACS applied materials & interfaces.

[7]  J. Madden,et al.  Electric field and vibration-assisted nanomolecule desorption and anti-biofouling for biosensor applications. , 2007, Colloids and surfaces. B, Biointerfaces.

[8]  C. Vahl,et al.  Cell adhesive and antifouling polyvinyl chloride surfaces via wet chemical modification. , 2012, Artificial organs.

[9]  Robert S. Marks,et al.  Chemiluminescent optical fiber immunosensor for the detection of IgM antibody to dengue virus in humans , 2009 .

[10]  Yung Chang,et al.  Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection. , 2012, Analytical biochemistry.

[11]  A. Guiseppi-Elie,et al.  Bio-smart hydrogels: co-joined molecular recognition and signal transduction in biosensor fabrication and drug delivery. , 2002, Biosensors & bioelectronics.

[12]  Hongwei Wang,et al.  Anti-fouling bioactive surfaces. , 2011, Acta biomaterialia.

[13]  D. Hobara,et al.  Preferential Adsorption of Horse Heart Cytochrome c on Nanometer-Scale Domains of a Phase-Separated Binary Self-Assembled Monolayer of 3-Mercaptopropionic Acid and 1-Hexadecanethiol on Au(111) , 2002 .

[14]  Yudong Zheng,et al.  Electrospun fibro-porous polyurethane coatings for implantable glucose biosensors. , 2013, Biomaterials.

[15]  R. F. Dutra,et al.  A disposable chitosan-modified carbon fiber electrode for dengue virus envelope protein detection. , 2012, Talanta.

[16]  A. Gopalan,et al.  One-step modification of various electrode surfaces using diazonium salt compounds and the application of this technology to electrochemical DNA (E-DNA) sensors , 2012 .

[17]  R. Suleiman,et al.  Current and emerging environmentally-friendly systems for fouling control in the marine environment. , 2013, Biotechnology advances.

[18]  S. Kiatkamjornwong,et al.  Development of a novel antifouling platform for biosensing probe immobilization from methacryloyloxyethyl phosphorylcholine-containing copolymer brushes. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[19]  Sook Mei Khor,et al.  A comparative study of the modification of gold and glassy carbon surfaces with mixed layers of in situ generated aryl diazonium compounds , 2010 .

[20]  Changsheng Liu,et al.  Development of mussel adhesive polypeptide mimics coating for in-situ inducing re-endothelialization of intravascular stent devices. , 2009, Biomaterials.

[21]  Zhongyi Jiang,et al.  Grafting short-chain amino acids onto membrane surfaces to resist protein fouling , 2011 .

[22]  Yuh-Chang Sun,et al.  Immunocapture couples with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid detection of type 1 dengue virus. , 2013, Journal of chromatography. A.

[23]  D. Bélanger,et al.  Metallic and bimetallic Cu/Pt species supported on carbon surfaces by means of substituted phenyl groups , 2007 .

[24]  D. Bélanger,et al.  Direct Modification of a Gold Electrode with Aminophenyl Groups by Electrochemical Reduction of in Situ Generated Aminophenyl Monodiazonium Cations , 2006 .

[25]  Peng Liu,et al.  Surface-initiated atom transfer radical polymerization (SI-ATRP) of styrene from chitosan particles , 2006 .

[26]  J. Gooding,et al.  The fabrication of stable gold nanoparticle-modified interfaces for electrochemistry. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[27]  Ronen Polsky,et al.  Electrically addressable diazonium-functionalized antibodies for multianalyte electrochemical sensor applications. , 2008, Biosensors & bioelectronics.

[28]  J. Gooding,et al.  Importance of the indium tin oxide substrate on the quality of self-assembled monolayers formed from organophosphonic acids. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[29]  C. Vieu,et al.  Arrays of nanoelectromechanical biosensors functionalized by microcontact printing , 2012, Nanotechnology.

[30]  C. Malitesta,et al.  X-ray photoelectron spectroscopy characterization of poly(2,3-diaminophenazine) films electrosynthesised on platinum , 2005 .

[31]  J. Gooding,et al.  Using supramolecular binding motifs to provide precise control over the ratio and distribution of species in multiple component films grafted on surfaces: demonstration using electrochemical assembly from aryl diazonium salts. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[32]  J. Gooding,et al.  Zwitterionic phenyl layers: finally, stable, anti-biofouling coatings that do not passivate electrodes. , 2013, ACS applied materials & interfaces.

[33]  Mohammed Zourob,et al.  Electrochemical immunosensor for the milk allergen β-lactoglobulin based on electrografting of organic film on graphene modified screen-printed carbon electrodes. , 2012, Biosensors & bioelectronics.

[34]  Guozhen Liu,et al.  Electrochemical impedance immunosensor based on gold nanoparticles and aryl diazonium salt functionalized gold electrodes for the detection of antibody. , 2011, Biosensors & bioelectronics.

[35]  D. Bélanger,et al.  The electrochemical grafting of a mixture of substituted phenyl groups at a glassy carbon electrode surface. , 2008, Chemphyschem : a European journal of chemical physics and physical chemistry.

[36]  D. Bélanger,et al.  Mixtures of functionalized aromatic groups generated from diazonium chemistry as templates towards bimetallic species supported on carbon electrode surfaces , 2012 .

[37]  Shuichi Takayama,et al.  Zwitterionic SAMs that Resist Nonspecific Adsorption of Protein from Aqueous Buffer. , 2001, Langmuir : the ACS journal of surfaces and colloids.

[38]  H. Zuilhof,et al.  Generic top-functionalization of patterned antifouling zwitterionic polymers on indium tin oxide. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[39]  M. Trau,et al.  An electrochemical immunosensor to minimize the nonspecific adsorption and to improve sensitivity of protein assays in human serum. , 2012, Biosensors & bioelectronics.

[40]  C. Combellas,et al.  Sterically hindered diazonium salts for the grafting of a monolayer on metals. , 2008, Journal of the American Chemical Society.