1. Orientation of llama antibodies strongly increases sensitivity of biosensors.
- Author
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Trilling AK, Hesselink T, van Houwelingen A, Cordewener JH, Jongsma MA, Schoffelen S, van Hest JC, Zuilhof H, and Beekwilder J
- Subjects
- Animals, Equipment Design, Equipment Failure Analysis, Protein Conformation, Reproducibility of Results, Sensitivity and Specificity, Antibodies, Viral immunology, Biosensing Techniques instrumentation, Camelids, New World immunology, Foot-and-Mouth Disease Virus immunology, Foot-and-Mouth Disease Virus isolation & purification, Immunoassay instrumentation, Surface Plasmon Resonance instrumentation
- Abstract
Sensitivity of biosensors depends on the orientation of bio-receptors on the sensor surface. The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups) to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
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