Rigid, specific, and discrete gold nanoparticle/antibody conjugates.
A general method of rigid, specific labeling of proteins with gold clusters has been devised. The method relies on the conjugation of a glutathione monolayer-protected gold cluster (MPC) with a single chain Fv antibody fragment (scFv), mutated to present an exposed cysteine residue. Efficient formation of a gold-thiolate bond between the MPC and scFv depends on activation of the gold cluster by chemical oxidation. Once formed, the MPC-scFv conjugate is treated with a reductant to quench cluster reactivity. The procedure has been performed with an MPC with an average Au(71) core and an scFv directed against a tetrameric protein, the influenza neuraminidase. A complex of the MPC-scFv conjugate with the neuraminidase was isolated, and the presence of four gold clusters was verified by cryoelectron microscopy.
Ackerson
Christopher J
CJ
Department of Structural Biology, Stanford University School of Medicine, California 94305, USA.
Jadzinsky
Pablo D
PD
Jensen
Grant J
GJ
Kornberg
Roger D
RD
D002852:Chromatography, Ion Exchange; D003545:Cysteine / Q000737:chemistry; D005978:Glutathione / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D018796:Immunoconjugates / Q000737:chemistry* / Q000302:isolation & purification; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D016297:Mutagenesis, Site-Directed; D049329:Nanostructures / Q000737:chemistry*; D009439:Neuraminidase / Q000737:chemistry; D010084:Oxidation-Reduction
D018796:Immunoconjugates; D007128:Immunoglobulin Fragments; D006046:Gold; D009439:Neuraminidase; D005978:Glutathione; D003545:Cysteine