Antibody-protein A conjugated quantum dots for multiplexed imaging of surface receptors in living cells.
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 F(c) (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.
Jin
Takashi
T
WPI Immunology Frontier Research Center, Osaka University, Yamada-oka 1-3, Suita, Osaka 565-0871, Japan. jin@fbs.osaka-u.ac.jp
Tiwari
Dhermendra K
DK
Tanaka
Shin-Ichi
S
Inouye
Yasushi
Y
Yoshizawa
Keiko
K
Watanabe
Tomonobu M
TM
D000906:Antibodies / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D045663:Quantum Dots; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013205:Staphylococcal Protein A / Q000378:metabolism*
D000906:Antibodies; D011956:Receptors, Cell Surface; D013205:Staphylococcal Protein A; D005978:Glutathione