Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays

Anal Chem. 2002 Feb 15;74(4):841-7. doi: 10.1021/ac010662m.

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).

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antibodies / chemistry*
  • Fluorescent Antibody Technique*
  • Fluorometry
  • Immunoglobulin G / chemistry
  • Indicators and Reagents
  • Luminescent Measurements
  • Molecular Sequence Data
  • Protein Engineering / methods*
  • Trinitrotoluene / chemistry

Substances

  • Antibodies
  • Immunoglobulin G
  • Indicators and Reagents
  • Trinitrotoluene