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Biosens Bioelectron ; 73: 100-107, 2015 Nov 15.
Article in English | MEDLINE | ID: mdl-26056953

ABSTRACT

Regeneration is a key goal in the design of immunosensors. In this study, we report the temperature-regulated interaction of N-isopropylacrylamide (PNIPAAm) functionalised cardiac troponin T (cTnT) with anti-cTnT. Covalently bonded PNIPAAm on an anti-cTnT bioelectrode showed on/off-switchability, regeneration capacity and temperature triggered sensitivity for cTnT. Above the lower critical solution temperature (LCST), PNIPAAm provides a liphophilic microenvironment with specific volume reduction at the bioelectrode surface, making available binding space for cTnT, and facilitating analyte recognition. Computational studies provide details about the structural changes occurring at the electrode above and below the LCST. Furthermore, free energies associated with the binding of cTnT with PNIPAAm at 25 (ΔGcoil=-6.0 Kcal/mole) and 37 °C (ΔGglobular=-41.0 kcal/mole) were calculated to elucidate the interaction and stability of the antigen-antibody complex. The responsiveness of such assemblies opens the way for miniaturised, smart immuno-technologies with 'built-in' programmable interactions of antigen-antibody upon receiving stimuli.


Subject(s)
Biosensing Techniques/methods , Troponin T/analysis , Acrylic Resins , Antibodies, Immobilized/chemistry , Antigen-Antibody Complex/chemistry , Electrochemical Techniques , Equipment Reuse , Humans , Immunoassay/methods , Models, Molecular , Molecular Dynamics Simulation , Protein Conformation , Temperature , Thermodynamics , Transition Temperature , Troponin T/immunology
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