Plant Research International, Wageningen, The Netherlands; Laboratory of Organic Chemistry, Wageningen University, Wageningen, The Netherlands.
Plant Research International, Wageningen, The Netherlands.
Biosens Bioelectron. 2014 Oct 15;60:130-6. doi: 10.1016/j.bios.2014.04.017. Epub 2014 Apr 18.
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.
生物传感器的灵敏度取决于传感器表面上生物受体的取向。本研究的目的是将生物受体组织在一种方式上，使它们的分析物结合位点暴露于分析物溶液中。针对口蹄疫病毒 (FMDV) 识别的 VHH 蛋白被用于制造生物传感器，并且在 VHH 中引入叠氮化物作为生物正交反应基团。通过将具有随机取向的 VHH（具有多个暴露的叠氮基团）的传感器与具有均匀取向的 VHH（只有一个叠氮基团）的传感器进行比较，来解决生物受体取向的重要性。使用点击化学，将暴露环辛炔的表面等离子体共振 (SPR) 芯片与叠氮功能化的 VHH 结构域反应。随机和均匀取向的生物受体之间的比较显示出生物传感器灵敏度提高了 800 倍。该技术可以增加对传染病（如口蹄疫病毒）的控制，因为其强烈增强的灵敏度可以促进早期诊断。
