Midland Blotting: A Rapid, Semi-Quantitative Method for Biosensor Surface CharacterizationJo V Rushworth*, Asif Ahmed and Paul A Millner
School of Biomedical Sciences, Faculty of Biological Science, University of Leeds, UK
- *Corresponding Author:
- Dr. Jo V Rushworth
School of Biomedical Sciences
Faculty of Biological Science
University of Leeds, UK
Tel: 00 44 113 34 37753
E-mail: [email protected]
Received Date: November 26, 2013; Accepted Date: December 19, 2013; Published Date: December 26, 2013
Citation: Rushworth JV, Ahmed A, Millner PA (2013) Midland Blotting: A Rapid, Semi-Quantitative Method for Biosensor Surface Characterization. J Biosens Bioelectron 4:146. doi: 10.4172/2155-6210.1000146
Copyright: © 2013 Rushworth JV, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Biosensor performance and readout are critically dependent upon sensor surface characteristics. It is vital that key steps in sensor construction, such as base layer polymer/Self-Assembled Monolayer (SAM) deposition and bioreceptor tethering, are controlled tightly in order to achieve high sensitivity, specificity and reproducibility. Here, we present a rapid, semi-quantitative method by which key biosensor surface features can be characterised using chemiluminescence. This technique, which we have termed midland blotting, permits the detection of biosensor surface components through the attachment of a HorseRadish Peroxidase (HRP)-conjugated reagent to the target of interest. Upon addition of luminol-based substrate, HRP generates a reagent which emits light when it decays. The light signal is proportional to the bound HRP on the sensor surface. We show here that midland blotting allows the measurement and validation of various important surface features including: (1) availability of functional groups on the polymer or SAM layer; (2) bioreceptor tethering and (3) analyte binding. Midland blotting is rapid, cost-effective and allows for much faster optimisation of biosensor surface design. This method also provides a simple way of troubleshooting and can explain sensor performance when combined with readout data. In this report, we have focussed on midland blotting for electrochemical immunosensors as a proof of concept, but this technique is readily applicable to all biosensor systems.