Conjugation of Luminescent Quantum Dots with Antibodies Using an Engineered Adaptor Protein To Provide New Reagents for Fluoroimmunoassays

Ellen R. Goldman,*† George P. Anderson,† Phan T. Tran,† Hedi Mattoussi,*‡ Paul T. Charles,† and J. Matthew Mauro†
Center for Bio/Molecular Science and Engineering and Division of Optical Sciences, U.S. Naval Research Laboratory, Washington, D.C. 20375
Anal. Chem., 2002, 74 (4), pp 841–847
DOI: 10.1021/ac010662m
Publication Date (Web): January 3, 2002
Copyright Not subject to U.S. Copyright. Published 2002 American Chemical Society
*

 Corresponding authors. E.R.G.:  (e-mail) erg@cbmse.nrl.navy.mil; (tel) 202-404-6052; (fax) 202-767-9594. H.M.:  (e-mail) hedimat@ccs.nrl.navy.mil; (tel) 202-767-9473.

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†

 Center for Bio/Molecular Science and Engineering.

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‡

 Division of Optical Sciences.

Abstract

We describe the preparation and characterization of bioinorganic conjugates made with highly luminescent semiconductor CdSe−ZnS core−shell quantum dots (QDs) and antibodies for use in fluoroimmunoassays. The conjugation strategy employs an engineered molecular adaptor protein, attached to the QDs via electrostatic/hydrophobic self-assembly, to link the inorganic fluorophore with antibodies. In this method, the number of antibodies conjugated to a single QD can be varied. In addition, we have developed a simple purification strategy based on mixed-composition conjugates of the molecular adaptor and a second two-domain protein that allows the use of affinity chromatography. QD−antibody conjugates were successfully used in fluoroimmunoassays for detection of both a protein toxin (staphylococcal enterotoxin B) and a small molecule (2,4,6-trinitrotoluene).

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History

  • Published In Issue February 15, 2002
  • Received for review June 14, 2001. Accepted October 9, 2001.

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