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Capping of CdSe–ZnS quantum dots with DHLA and subsequent conjugation with proteins

Nature Protocols volume 1, pages 1258–1266 (2006)Cite this article

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Abstract

We provide a detailed protocol for designing water-soluble CdSe–ZnS quantum dots (QDs) based on cap exchange of the native hydrophobic shell with dihydrolipoic acid (DHLA) ligands, and the preparation of functional QD bioconjugates for use in immunoassays. Our conjugation strategy is based on non-covalent self-assembly between DHLA-capped QDs and protein appended with either an electrostatic attachment domain (namely, the basic leucine zipper) or a polyhistidine tag. These bioconjugates combine the properties of the QD and attached biomolecule to create structures with desirable luminescent and biologically specific properties. This method also allows the preparation of mixed surface conjugates, which results in the conjugates gaining multiple biological activities. Conjugation of DHLA-capped QDs to maltose binding protein (MBP), the immunoglobulin-G-binding β2 domain of streptococcal protein G (PG) and avidin will be described. MBP and PG were modified by genetic fusion with either a charged leucine zipper or a polyhistidine interaction domain.

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Figure 1: Schematic representation of a typical QD bioconjugate.
Figure 2: Multiplexed toxin analysis using QD–antibody conjugates.
Figure 3: Results from a competition assay using QD–anti-TNT scFv conjugate to detect soluble TNT.

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  • 30 November 2006

    In the version of this article initially published online, the article’s page numbers should have been 1258–1266. This error has been corrected in the PDF version of the article.

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Acknowledgements

This work was supported by grants from the Office of Naval Research (ONR) and DARPA. A.R.C. was supported by a National Research Council fellowship.

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Authors and Affiliations

  1. Optical Sciences Division, US Naval Research Laboratory, Code 5611, 4555 Overlook Avenue SW, Washington, 20375, DC, USA

    Aaron R. Clapp & Hedi Mattoussi

  2. Center for Bio/Molecular Science and Engineering, Code 6900, 4555 Overlook Avenue SW, Washington, 20375, DC, USA

    Ellen R. Goldman

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  1. Aaron R. Clapp
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Correspondence to Hedi Mattoussi.

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Clapp, A., Goldman, E. & Mattoussi, H. Capping of CdSe–ZnS quantum dots with DHLA and subsequent conjugation with proteins. Nat Protoc 1, 1258–1266 (2006). https://doi.org/10.1038/nprot.2006.184

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