A new application of scanning electrochemical microscopy for the label-free interrogation of antibody-antigen interactions.
| dc.contributor.author | Holmes, Joanne L. | - |
| dc.contributor.author | Davis, Frank | - |
| dc.contributor.author | Collyer, Stuart D. | - |
| dc.contributor.author | Higson, Seamus P. J. | - |
| dc.date.accessioned | 2011-12-01T23:00:46Z | |
| dc.date.available | 2011-12-01T23:00:46Z | |
| dc.date.issued | 2011-03-18T00:00:00Z | - |
| dc.description.abstract | Within this work we present a 'proof of principle' study for the use of scanning electrochemical microscopy (SECM) to detect and image biomolecular interactions in a label-free assay as a potential alternative to current fluorescence techniques. Screen-printed carbon electrodes were used as the substrate for the deposition of a dotted array, where the dots consist of biotinylated polyethyleneimine. These were then further derivatised, first with neutravidin and then with a biotinylated antibody to the protein neuron specific enolase (NSE). SECM using a ferrocene carboxylic acid mediator showed clear differences between the array and the surrounding unmodified carbon. Imaging of the arrays before and following exposure to various concentrations of the antigen showed clear evidence for specific binding of the NSE antigen to the antibody derivatised dots. Non-specific binding was quantified. Control experiments with other proteins showed only non-specific binding across the whole of the substrate, thereby confirming that specific binding does occur between the antibody and antigen at the surface of the dots. Binding of the antigen was accompanied by a measured increase in current response, which may be explained in terms of protein electrostatic interaction and hydrophobic interactions to the mediator, thereby increasing the localised mediator flux. A calibration curve was obtained between 500 fg mL(-1) to 200 pg mL(-1) NSE which demonstrated a logarithmic relationship between the current change upon binding and antigen concentration without the need for any labelling of the substrate. | en_UK |
| dc.identifier.citation | Joanne L. Holmes, Frank Davis, Stuart D. Collyer, Séamus P.J. Higson, A new application of scanning electrochemical microscopy for the label-free interrogation of antibody-antigen interactions, Analytica Chimica Acta, Volume 689, Issue 2, 18 March 2011, Pages 206-211. | |
| dc.identifier.issn | 0003-2670 | - |
| dc.identifier.uri | http://dx.doi.org/10.1016/j.aca.2011.01.033 | - |
| dc.identifier.uri | http://dspace.lib.cranfield.ac.uk/handle/1826/6768 | |
| dc.publisher | Elsevier Science B.V., Amsterdam. | en_UK |
| dc.rights | NOTICE: this is the author’s version of a work that was accepted for publication in Analytica Chimica Acta. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Analytica Chimica Acta, VOL 689, ISSUE 2, (2011) DOI:10.1016/j.aca.2011.01.033 | |
| dc.title | A new application of scanning electrochemical microscopy for the label-free interrogation of antibody-antigen interactions. | en_UK |
| dc.type | Article | - |