Abstract
Nanoparticle-based biodetection routinely employs monoclonal antibodies (mAbs) for targeting. However, the large size of mAbs limits the number of ligands per nanoparticle and severely restricts the bioavailability and distribution of these probes in a sample. Furthermore, conventional conjugation techniques provide nanoprobes with irregular orientation of mAbs on the nanoparticle surface and often provoke mAb unfolding. Here, we describe a protocol for engineering a new generation of ultrasmall diagnostic nanoprobes through oriented conjugation of semiconductor quantum dots (QDs) with 13 kDa single-domain antibodies (sdAbs) derived from llama immunoglobulin G (IgG). The sdAbs are conjugated with QDs in a highly oriented manner via an additional cysteine residue specifically integrated into the sdAb C-terminus. The resultant nanoprobes are <12 nm in diameter, ten times smaller in volume compared to the known alternatives. They have been proved highly efficient in flow cytometry and immunuhistochemical diagnostics. This approach can be easily extended to other semiconductor and plasmonic nanoparticles.
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Acknowledgements
This study was partly supported by the Ministry of Education and Science of the Russian Federation (grant 11.G34.31.0050), by the European Commission through the FP7 Cooperation project NAMDIATREAM (grant NMP-2009-4.0-3-246479) and by the programs HYNNOV and Nano’Mat of the Champagne-Ardenne region, the DRRT Champagne-Ardenne, and the FEDER (France). I.N. acknowledges support of the Russian Science Foundation.
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Brazhnik, K., Nabiev, I., Sukhanova, A. (2014). Oriented Conjugation of Single-Domain Antibodies and Quantum Dots. In: Fontes, A., Santos, B. (eds) Quantum Dots: Applications in Biology. Methods in Molecular Biology, vol 1199. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1280-3_10
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DOI: https://doi.org/10.1007/978-1-4939-1280-3_10
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