Issue 11, 2010

AntibodyProteinA conjugated quantum dots for multiplexed imaging of surface receptors in living cells

Abstract

To use quantum dots (QDs) as fluorescent probes for receptor imaging, QD surface should be modified with biomolecules such as antibodies, peptides, carbohydrates, and small-molecule ligands for receptors. Among these QDs, antibody conjugated QDs are the most promising fluorescent probes. There are many kinds of coupling reactions that can be used for preparing antibody conjugated QDs. Most of the antibody coupling reactions, however, are non-selective and time-consuming. In this paper, we report a facile method for preparing antibody conjugated QDs for surface receptor imaging. We used ProteinA as an adaptor protein for binding of antibody to QDs. By using ProteinA conjugated QDs, various types of antibodies are easily attached to the surface of the QDs via non-covalent binding between the Fc (fragment crystallization) region of antibody and ProteinA. To show the utility of ProteinA conjugated QDs, HER2 (anti-human epidermal growth factor receptor 2) in KPL-4 human breast cancer cells were stained by using anti-HER2 antibody conjugated ProteinA–QDs. In addition, multiplexed imaging of HER2 and CXCR4 (chemokine receptor) in the KPL-4 cells was performed. The result showed that CXCR4 receptors coexist with HER2 receptors in the membrane surface of KPL-4 cells. ProteinA mediated antibody conjugation to QDs is very useful to prepare fluorescent probes for multiplexed imaging of surface receptors in living cells.

Graphical abstract: Antibody–ProteinA conjugated quantum dots for multiplexed imaging of surface receptors in living cells

Supplementary files

Article information

Article type
Paper
Submitted
18 Jun 2010
Accepted
09 Aug 2010
First published
10 Sep 2010

Mol. BioSyst., 2010,6, 2325-2331

Antibody ProteinA conjugated quantum dots for multiplexed imaging of surface receptors in living cells

T. Jin, D. K. Tiwari, S. Tanaka, Y. Inouye, K. Yoshizawa and T. M. Watanabe, Mol. BioSyst., 2010, 6, 2325 DOI: 10.1039/C0MB00056F

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