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Sensors 2013, 13(4), 5273-5285; doi:10.3390/s130405273

Development of Robust and Standardized Cantilever Sensors Based on Biotin/Neutravidin Coupling for Antibody Detection

Swiss Nano Institute, University of Basel, Klingelbergstrasse 82, 4056 Basel, Switzerland
Concentris GmbH, Davidsbodenstrasse 63, 4056 Basel, Switzerland
Author to whom correspondence should be addressed.
Received: 14 March 2013 / Revised: 10 April 2013 / Accepted: 15 April 2013 / Published: 19 April 2013
(This article belongs to the Section Biosensors)
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A cantilever-based protein biosensor has been developed providing a customizable multilayer platform for the detection of antibodies. It consists of a biotin-terminated PEG layer pre-functionalized on the gold-coated cantilever surface, onto which NeutrAvidin is adsorbed through biotin/NeutrAvidin specific binding. NeutrAvidin is used as a bridge layer between the biotin-coated surface and the biotinylated biomolecules, such as biotinylated bovine serum albumin (biotinylated BSA), forming a multilayer sensor for direct antibody capture. The cantilever biosensor has been successfully applied to the detection of mouse anti-BSA (m-IgG) and sheep anti-BSA(s-IgG) antibodies. As expected, the average differential surface stress signals of about 5.7 ± 0.8 ´ 10−3 N/m are very similar for BSA/m-IgG and BSA/s-IgG binding, i.e., they are independent of the origin of the antibody. A statistic evaluation of 112 response curves confirms that the multilayer protein cantilever biosensor shows high reproducibility. As a control test, a biotinylated maltose binding protein was used for detecting specificity of IgG, the result shows a signal of bBSA layer in response to antibody is 5.8 ´ 10−3 N/m compared to bMBP. The pre-functionalized biotin/PEG cantilever surface is found to show a long shelf-life of at least 40 days and retains its responsivity of above 70% of the signal when stored in PBS buffer at 4 °C. The protein cantilever biosensor represents a rapid, label-free, sensitive and reliable detection technique for a real-time protein assay.
Keywords: cantilever; biosensor; protein; multilayer; NeutrAvidin; biotin cantilever; biosensor; protein; multilayer; NeutrAvidin; biotin

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Zhang, J.; Lang, H.P.; Battiston, F.; Backmann, N.; Huber, F.; Gerber, C. Development of Robust and Standardized Cantilever Sensors Based on Biotin/Neutravidin Coupling for Antibody Detection. Sensors 2013, 13, 5273-5285.

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