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Orientation of Llama Antibodies Strongly Increases Sensitivity of Biosensors by Anke K. Trilling; Thamara Hesselink; Adèle van Houwelingen; Jan H.G. Cordewener; Maarten A. Jongsma; Sanne Schoffelen; Jan C.M. van Hest; Han Zuilhof; Jules Beekwilder is a Engineering article available to read on EtoBox.

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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.

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Author

Anke K. Trilling; Thamara Hesselink; Adèle van Houwelingen; Jan H.G. Cordewener; Maarten A. Jongsma; Sanne Schoffelen; Jan C.M. van Hest; Han Zuilhof; Jules Beekwilder

Publisher

Elsevier BV

Published

2014

Language

EN

Field

Engineering (Physical Sciences)