Abstract
Luminescent colloidal semiconductor nanocrystals (quantum dots) are robust inorganic fluorophores that have the potential to circumvent some of the functional limitations encountered by organic dyes in sensing and biotechnological applications. Quantum dots exhibit size-dependent tunable, narrow fluorescence emission spectra that span the visible spectrum and have broad absorption spectra. This allows simultaneous excitation of several particle sizes at a single wavelength with emission at multiple wavelengths. Quantum dots also provide a high-resistance threshold to chemical degradation and photodegradation. We have developed a conjugation strategy for the attachment of antibodies to quantum dots based on electrostatic interactions between negatively charged dihydrolipoic acid (DHLA)-capped CdSe-ZnS core-shell quantum dots and positively charged proteins (natural or engineered) that serve to bridge the quantum dot and antibody. This chapter details the materials and methods for synthesis of the DHLA-capped CdSe-ZnS core-shell quantum dots, the construction and preparation of recombinant proteins, the conjugation of antibodies to quantum dots, and the use of antibody-coated quantum dots in a fluoroimmunoassay.
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Acknowledgments
We thank the Office of the Naval Research for research support. The views, opinions, and/or findings described in this chapter are those of the authors and should not be construed as official Department of the Navy positions, policies, or decisions.
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Goldman, E.R., Mattoussi, H., Anderson, G.P., Medintz, I.L., Mauro, J.M. (2005). Fluoroimmunoassays Using Antibody-Conjugated Quantum Dots. In: Rosenthal, S.J., Wright, D.W. (eds) NanoBiotechnology Protocols. Methods in Molecular Biology™, vol 303. Humana Press. https://doi.org/10.1385/1-59259-901-X:019
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DOI: https://doi.org/10.1385/1-59259-901-X:019
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