A novel biochip technology for detection of explosives – TNT: Synthesis, characterisation and application

This contribution describes the synthesis, characterisation and evaluation of a novel biochip technology for the detection of the explosive substance 2,4,6-trinitrotoluene (TNT). Two types of thiols are self-assembled to produce the biochip on gold, namely oligo(ethylene glycol) (OEG)-alkyl thiols terminated with a hydroxyl group and a TNT-analogue (2,4-dinitrobenzene), respectively. Three different TNT-analogues are mixed in various proportions with hydroxyl-terminated OEG-thiols to obtain highly selective and sensitive biochips with a low non-specific binding. The produced self-assembled monolayers (SAMs) are thoroughly characterised with null ellipsometry, contact angle goniometry, infrared reflection absorption spectroscopy (IRAS) and X-ray photoelectron spectroscopy (XPS) and they all meet high standards in terms of molecular conformation, packing and orientation. The biochip is designed to function as a platform for a competitive label-free immunoassay and two real-time transducers – surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) – are used to monitor the dissociation of on-line immobilised monoclonal antibodies produced against TNT. The three TNT-analogues are all potential candidates for the development of a functional biochip, though one of them displayed superior properties in terms of shorter recovery/stabilisation time after antibody immobilisation and a better response/loading capacity ratio. This is particularly evident when using low antigen (TNT-analogue) content in the mixed SAM.

[1]  P. Tengvall,et al.  Protein adsorption to oligo(ethylene glycol) self-assembled monolayers: Experiments with fibrinogen, heparinized plasma, and serum , 2001, Journal of biomaterials science. Polymer edition.

[2]  B. Kasemo,et al.  Variations in coupled water, viscoelastic properties, and film thickness of a Mefp-1 protein film during adsorption and cross-linking: a quartz crystal microbalance with dissipation monitoring, ellipsometry, and surface plasmon resonance study. , 2001, Analytical chemistry.

[3]  Raphael Lavi,et al.  Application of a unique scheme for remote detection of explosives , 2002 .

[4]  M. Oda,et al.  Reevaluation of stoichiometry and affinity/avidity in interactions between anti-hapten antibodies and mono- or multi-valent antigens. , 2000, Molecular immunology.

[5]  Petr Skládal,et al.  Improved direct piezoelectric biosensors operating in liquid solution for the competitive label-free immunoassay of 2,4-dichlorophenoxyacetic acid , 1997 .

[6]  B. Liedberg,et al.  Self-Assembled Monolayers of Oligo(ethylene glycol)-Terminated and Amide Group Containing Alkanethiolates on Gold , 1999 .

[7]  Paul T. Charlesa,et al.  Microcapillary reversed-displacement immunosensor for trace level detection of TNT in seawater , 2004 .

[8]  S. Kurosawa,et al.  Quartz crystal microbalance for the detection of microgram quantities of human serum albumin: relationship between the frequency change and the mass of protein adsorbed. , 1993, Analytical chemistry.

[9]  J. A. Evans,et al.  QCM response to solvated, tethered macromolecules. , 1998, Analytical chemistry.

[10]  Ralph G. Nuzzo,et al.  ADSORPTION OF BIFUNCTIONAL ORGANIC DISULFIDES ON GOLD SURFACES , 1983 .

[11]  Robert Wilson,et al.  Electrochemiluminescence enzyme immunoassays for TNT and pentaerythritol tetranitrate. , 2003, Analytical chemistry.

[12]  G. Whitesides,et al.  Self-assembled organic monolayers: model systems for studying adsorption of proteins at surfaces , 1991, Science.

[13]  B. Liedberg,et al.  Synthesis of a series of oligo(ethylene glycol)-terminated alkanethiol amides designed to address structure and stability of biosensing interfaces. , 2001, The Journal of organic chemistry.

[14]  Frances S. Ligler,et al.  Immobilized biomolecules in analysis : a practical approach , 1998 .

[15]  D. Allara,et al.  Spontaneously organized molecular assemblies. 4. Structural characterization of n-alkyl thiol monolayers on gold by optical ellipsometry, infrared spectroscopy, and electrochemistry , 1987 .

[16]  B. Swanson,et al.  Molecular recognition and self-assembled polymer films for vapor phase detection of explosives. , 2001, Talanta.

[17]  Caruso,et al.  Characterization of Ferritin Adsorption onto Gold , 1997, Journal of colloid and interface science.

[18]  Timothy A. Springer,et al.  Hybridoma Technology in the Biosciences and Medicine , 2012, Springer US.

[19]  Y. Fung,et al.  Study of adsorption behavior of bilirubin on human-albumin monolayer using a quartz crystal microbalance. , 2002, Journal of colloid and interface science.

[20]  B. Liedberg,et al.  Temperature-Driven Phase Transitions in Oligo(ethylene glycol)-terminated Self-Assembled Monolayers , 2000 .

[21]  W. Göpel,et al.  Interface analysis in biosensor design. , 1995, Biosensors & bioelectronics.

[22]  B. Liedberg,et al.  Functionalized Surfaces of Mixed Alkanethiols on Gold as a Platform for Oligonucleotide Microarrays , 2002 .

[23]  B. Liedberg,et al.  Thermal stability of self-assembled monolayers : Influence of lateral hydrogen bonding , 2002 .

[24]  H. Ringsdorf,et al.  Biotin-functionalized self-assembled monolayers on gold: surface plasmon optical studies of specific recognition reactions , 1991 .

[25]  George M. Whitesides,et al.  Biospecific Binding of Carbonic Anhydrase to Mixed SAMs Presenting Benzenesulfonamide Ligands: A Model System for Studying Lateral Steric Effects , 1999 .

[26]  M. Andersson,et al.  Synthesis and Self-Assembly of Globotriose Derivatives: A Model System for Studies of Carbohydrate−Protein Interactions , 2002 .

[27]  George M. Whitesides,et al.  Adsorption of Proteins to Hydrophobic Sites on Mixed Self-Assembled Monolayers† , 2003 .

[28]  George M. Whitesides,et al.  Molecular Conformation in Oligo(ethylene glycol)-Terminated Self-Assembled Monolayers on Gold and Silver Surfaces Determines Their Ability To Resist Protein Adsorption , 1998 .

[29]  G. Lubrano,et al.  A quartz crystal microbalance displacement assay for Listeria monocytogenes , 1996 .

[30]  A. Ulman,et al.  Ultrathin organic films: From Langmuir-Blodgett to self assembly , 1991 .

[31]  G. Whitesides,et al.  A Survey of Structure−Property Relationships of Surfaces that Resist the Adsorption of Protein , 2001 .

[32]  A W Kusterbeck,et al.  Explosives detection in soil using a field-portable continuous flow immunosensor. , 2001, Journal of hazardous materials.

[33]  J. Halámek,et al.  Highly sensitive detection of cocaine using a piezoelectric immunosensor. , 2002, Biosensors & bioelectronics.

[34]  B. Liedberg,et al.  Influence of specific intermolecular interactions on the self-assembly and phase behavior of oligo(ethylene glycol)-terminated alkanethiolates on gold , 2001 .

[35]  M. Grunze,et al.  Factors that determine the protein resistance of oligoether self-assembled monolayers --internal hydrophilicity, terminal hydrophilicity, and lateral packing density. , 2003, Journal of the American Chemical Society.

[36]  G. Whitesides,et al.  Self-Assembled Monolayers That Resist the Adsorption of Proteins and the Adhesion of Bacterial and Mammalian Cells , 2001 .

[37]  Frances S. Ligler,et al.  Multi-analyte explosive detection using a fiber optic biosensor , 1999 .

[38]  Joseph Wang,et al.  Carbon nanotube-modified glassy carbon electrode for adsorptive stripping voltammetric detection of ultratrace levels of 2,4,6-trinitrotoluene , 2004 .

[39]  J. Matthew Mauro,et al.  Analysis of aqueous 2,4,6-trinitrotoluene (TNT) using a fluorescent displacement immunoassay , 2003, Analytical and bioanalytical chemistry.

[40]  A. Ulman,et al.  Formation and Structure of Self-Assembled Monolayers. , 1996, Chemical reviews.

[41]  G. Sauerbrey,et al.  Use of quartz vibration for weighing thin films on a microbalance , 1959 .

[42]  Garry A. Rechnitz,et al.  Flow injection immunosensing of polycyclic aromatic hydrocarbons with a quartz crystal microbalance , 1999 .

[43]  G. Whitesides,et al.  Surface Plasmon Resonance Permits in Situ Measurement of Protein Adsorption on Self-Assembled Monolayers of Alkanethiolates on Gold , 1995 .

[44]  Richard A Mathies,et al.  Homogeneous immunoassay for detection of TNT and its analogues on a microfabricated capillary electrophoresis chip. , 2003, Analytical chemistry.

[45]  G. Sauerbrey.  Verwendung von Schwingquarzen zur Wägung dünner Schichten und zur Mikrowägung , 1959 .

[46]  W. Göpel,et al.  Self-assembled monolayers for chemical sensors: molecular recognition by immobilized supramolecular structure , 1996 .