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

Sensors

Jiayun Zhang²  Hans Peter Lang²  Felice Battiston¹  Natalija Backmann²  Francois Huber² 

[1] Concentris GmbH, Davidsbodenstrasse 63, 4056 Basel, Switzerland; E-Mail:;Swiss Nano Institute, University of Basel, Klingelbergstrasse 82, 4056 Basel, Switzerland; E-Mails:

关键词: cantilever;    biosensor;    protein;    multilayer;    NeutrAvidin;    biotin;   

DOI  :  10.3390/s130405273

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland.