Probing functionalized gold colloids for single particle tracking experiments.
Functionalized submicroscopic particles are currently used to label proteins or lipids at the surface of living cells for single particle tracking experiments. In many cases, it can be of crucial importance for the particle to be anchored to a single molecule. We have addressed this question for the labeling at the plasma membrane of NRK cells of the mu-opioid receptor bearing a T7 epitope at the N-terminus. Using biophysical methods we were able to prepare quasi-monovalent anti-T7 antibody conjugated gold colloids (40 nm diameter) leading to stable and specific binding to the receptor. The rational method, we report here, can be extended to design customized probes for the labeling of various tagged molecules.
Daumas
Frédéric
F
Institut de Pharmacologie et Biologie Structurale, CNRS UMR 5089, 205, route de Narbonne, Toulouse cedex 31077, France.
Mazarguil
Honoré
H
Millot
Claire
C
Lopez
André
A
Salomé
Laurence
L
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure; D003102:Colloids; D006046:Gold / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007128:Immunoglobulin Fragments; D007526:Isoelectric Point; D007700:Kinetics; D008055:Lipids; D008853:Microscopy / Q000379:methods*; D051381:Rats; D017450:Receptors, Opioid, mu / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology; D020349:Surface Plasmon Resonance; D013997:Time Factors; D014162:Transfection
D003102:Colloids; D007128:Immunoglobulin Fragments; D008055:Lipids; D017450:Receptors, Opioid, mu; D006046:Gold; D012964:Sodium