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A Label-free Immunosensor Array Using Single-chain Antibody Fragments.
N. Backmann, C. Zahnd, F. Huber, A. Bietsch, A. Plückthun, Hans-Peter Lang, H. Guentherodt, M. Hegner, C. Gerber
Published 2005 · Medicine, Chemistry
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We report a microcantilever-based immunosensor operated in static deflection mode with a performance comparable with surface plasmon resonance, using single-chain Fv (scFv) antibody fragments as receptor molecules. As a model system scFv fragments with specificity to two different antigens were applied. We introduced a cysteine residue at the C terminus of each scFv construct to allow covalent attachment to gold-coated sensor interfaces in directed orientation. Application of an array enabled simultaneous deflection measurements of sensing and reference cantilevers. The differential deflection signal revealed specific antigen binding and was proportional to the antigen concentration in solution. Using small, oriented scFv fragments as receptor molecules we increased the sensitivity of microcantilevers to approximately 1 nM.
This paper references
Orientation of Adsorbed Antibodies on Charged Surfaces by Computer Simulation Based on a United-Residue Model
Jian Zhou (2003)
Determination of rate and equilibrium binding constants for macromolecular interactions using surface plasmon resonance: use of nonlinear least squares analysis methods.
D. O'shannessy (1993)
Cancer nanotechnology: opportunities and challenges
M. Ferrari (2005)
Bioassay of prostate-specific antigen (PSA) using microcantilevers
G. Wu (2001)
Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system.
A. Krebber (1997)
Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display
J. Hanes (2000)
Multiple label-free biodetection and quantitative DNA-binding assays on a nanomechanical cantilever array
R. McKendry (2002)
Gold Nano-Structures for Transduction of Biomolecular Interactions into Micrometer Scale Movements
N. Lavrik (2001)
Designing repeat proteins: well-expressed, soluble and stable proteins from combinatorial libraries of consensus ankyrin repeat proteins.
H. Binz (2003)
Rapid functionalization of cantilever array sensors by inkjet printing
A. Bietsch (2004)
High‐density immobilization of an antibody fragment to a carboxymethylated dextran‐linked biosensor surface
S. Howell (1998)
Antigen binding forces of individually addressed single-chain Fv antibody molecules.
R. Ros (1998)
Microarrays based on affinity‐tagged single‐chain Fv antibodies: Sensitive detection of analyte in complex proteomes
C. Wingren (2005)
Assembling of engineered IgG-binding protein on gold surface for highly oriented antibody immobilization.
S. Kanno (2000)
Characterization of ultrathin poly(ethylene glycol) monolayers on silicon substrates
A. Papra (2001)
Label-free protein assay based on a nanomechanical cantilever array
Y. Arntz (2003)
Translating biomolecular recognition into nanomechanics.
J. Fritz (2000)
Managing heat for electronics
P. Schelling (2005)
Enantioselective sensors based on antibody-mediated nanomechanics.
P. Dutta (2003)
Systems Biology and New Technologies Enable Predictive and Preventative Medicine
Leroy E Hood (2004)
Wafer-scale microdevice transfer/interconnect: its application in an AFM-based data-storage system
M. Despont (2004)
Covalent immobilization of native biomolecules onto Au(111) via N-hydroxysuccinimide ester functionalized self-assembled monolayers for scanning probe microscopy.
P. Wagner (1996)
Antibody microarrays: promises and problems.
W. Kusnezow (2002)
Evaluation of a high-affinity QCM immunosensor using antibody fragmentation and 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer.
S. Kurosawa (2004)
Micromechanical detection of proteins using aptamer-based receptor molecules.
C. Savran (2004)
Ribosome display efficiently selects and evolves high-affinity antibodies in vitro from immune libraries.
J. Hanes (1998)
Directed in Vitro Evolution and Crystallographic Analysis of a Peptide-binding Single Chain Antibody Fragment (scFv) with Low Picomolar Affinity*
C. Zahnd (2004)
Micromechanical cantilever-based biosensors
R. Raiteri (2001)
Oriented immobilization of antibodies onto the gold surfaces via their native thiol groups.
A. A. Karyakin (2000)
Influence of surfactants and antibody immobilization strategy on reducing nonspecific protein interactions for molecular recognition force microscopy.
K. Brogan (2004)
Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides.
A. Knappik (2000)
Prostate-specific antigen immunosensing based on mixed self-assembled monolayers, camel antibodies and colloidal gold enhanced sandwich assays.
Lieven Huang (2005)
Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries
E. Söderlind (2000)
High-affinity binders selected from designed ankyrin repeat protein libraries
H Kaspar Binz (2004)
Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins
E. Stenberg (1991)
In vitro selection and evolution of functional proteins by using ribosome display.
J. Hanes (1997)
Unbinding forces of single antibody-antigen complexes correlate with their thermal dissociation rates.
F. Schwesinger (2000)
Cantilever transducers as a platform for chemical and biological sensors
N. Lavrik (2004)
Optimizing antibody immobilization strategies for the construction of protein microarrays.
Paul Peluso (2003)
Antibodies from Escherichia coli
A. Plückthun (1990)
Measuring Surface-Induced Conformational Changes in Proteins
A. Moulin (1999)
Characterisation of an antibody coated microcantilever as a potential immuno-based biosensor.
C. Grogan (2002)
Real time, high resolution studies of protein adsorption and structure at the solid liquid interface using dual polarization interferometry
N. Freeman (2004)
Making antibodies by phage display technology.
G. Winter (1994)
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Antibody microarrays: current status and key technological advances.
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Comparative advantages of mechanical biosensors.
J. Arlett (2011)
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Ming-Liang Lai (2015)
Survey of the year 2005 commercial optical biosensor literature
R. Rich (2006)
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Y. Nemirovsky (2008)
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Genetic engineering of recombinant anti-mycolic acid antibody fragments for use in tuberculosis diagnostics
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Micro- and nanocantilever devices and systems for biomolecule detection.
K. S. Hwang (2009)
Quantitative, Label-Free Detection of the Aggregation of α-Synuclein Using Microcantilever Arrays Operated in a Liquid Environment
Jason Jensen (2012)
Succinic anhydride functionalized microcantilevers for protein immobilization and quantification
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Deflection cantilever detection of interferon gamma
R. V. D. Hurk (2013)
Microcantilever biosensors: probing biomolecular interactions at the nanoscale
D. Liu (2011)
A novel fabrication technique for free-hanging homogeneous polymeric cantilever waveguides
M. Nordström (2008)
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Stanley J. Ness (2012)
Quantitative time-resolved measurement of membrane protein-ligand interactions using microcantilever array sensors.
T. Braun (2009)
Use of biospecific reactions for the design of high-sensitivity biosensors based on nanomechanical cantilever systems
P. Gorelkin (2010)
Observation of nanoscale dynamics in cantilever sensor arrays
J. Reed (2006)
Antibody-based protein detection using piezoresistive cantilever arrays
V. Dauksaite (2007)
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