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|Document Type: ||Article|
|Title: ||Labeless and reversible immunosensor assay based upon an electrochemical
|Authors: ||Grant, Sarah|
Pritchard, Jeanette A.
Law, Karen A.
Higson, Seamus P. J.
Gibson, Timothy D.
|Issue Date: ||2003|
|Citation: ||Sarah Grant, Frank Davis, Jeanette A. Pritchard, Karen A. Law, Seamus P. J. Higson, and Timothy D. Gibson, Labeless and reversible immunosensor assay based upon an electrochemical current-transient protocol, Analytica Chimica Acta, Volume 495, Issues 1-2, 24 October 2003, Pages 21-32|
|Abstract: ||A novel labeless and reversible immunoassay based upon an electrochemical
current-transient protocol is reported which offers many advantages in
comparison to classical immuno-biochemical analyses in terms of simplicity,
speed of response, reusability and possibility of multiple determinations.
Conducting polypyrrole films containing antibodies against 1) Bovine Serum
Albumin (BSA) and 2) Digoxin were deposited on the surface of platinum
electrodes to produce conductive affinity matrices having clearly defined
binding characteristics. The deposition process has been investigated using 125I
labelled anti-digoxin to determine optimal fabrication protocols. Antibody
integrity and activity, together with non-specific binding of antigen on the
conducting matrix have also been investigated using tritiated digoxin to probe
polypyrrole/anti-digoxin films. Amperometric responses to digoxin were recorded
in flow conditions using these films, but the technique was limited in use
mainly due to baseline instability. Anti-BSA - polypyrrole matrices were
investigated in more detail in both flow and quiescent conditions. No observable
response was found in flow conditions, however under quiescent conditions (in
non-stirred batch cell), anti-BSA – polypyrrole films have been demonstrated to
function as novel quantitative chronoamperometric immuno-biosensors when
interrogated using a pulsed potential waveform. The behaviour of the electrodes
showed that the antibody/antigen binding and/or interaction process underlying
the response observed was reversible in nature, indicating that the electrodes
could be used for multiple sensing protocols. Calibration profiles for BSA
demonstrated linearity for a concentration range of 0-50 ppm but tended towards
a plateau at higher concentrations. Factors relating to replicate sensor
production, sample measurement and reproducibility are discusse|
|Appears in Collections:||Staff publications - Cranfield University at Silsoe|
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