title: One-Step Protein Conjugation to Upconversion Nanoparticles.
abstract: The emerging upconversion nanoparticles offer a fascinating library of ultrasensitive luminescent probes for a range of biotechnology applications from biomarker discovery to single molecule tracking, early disease diagnosis, deep tissue imaging, and drug delivery and therapies. The effective bioconjugation of inorganic nanoparticles to the molecule-specific proteins, free of agglomeration, nonspecific binding, or biomolecule deactivation, is crucial for molecular recognition of target molecules or cells. The current available protocols require multiple steps which can lead to low probe stability, specificity, and reproducibility. Here we report a simple and rapid protein bioconjugation method based on a one-step ligand exchange using the DNAs as the linker. Our method benefits from the robust DNA-protein conjugates as well as from multiple ions binding capability. Protein can be preconjugated via an amino group at the 3' end of a synthetic DNA molecule, so that the 5' end phosphoric acid group and multiple phosphate oxygen atoms in the phosphodiester bonds are exposed to replace the oleic acid ligands on the surface of upconversion nanoparticles due to their stronger chelating capability to lanthanides. We demonstrated that our method can efficiently pull out the upconversion nanoparticles from organic solvent into an aqueous phase. The upconversion nanoparticles then become hydrophilic, stable, and specific biomolecules recognition. This allows us to successfully functionalize the upconversion nanoparticles with horseradish peroxidise (HRP) for catalytic colorimetric assay and for streptavidin (SA)-biotin immunoassays.
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.affiliation: Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology, Sydney, NSW 2007, Australia.
authors.forename: Yinghui
authors.initials: Y
authors.lastname: Chen
authors.forename: Deming
authors.initials: D
authors.lastname: Liu
authors.affiliation: Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology, Sydney, NSW 2007, Australia.
authors.forename: Wei
authors.initials: W
authors.lastname: Ren
authors.forename: Yiqing
authors.initials: Y
authors.lastname: Lu
authors.forename: Yu
authors.initials: Y
authors.lastname: Shi
authors.forename: James
authors.initials: J
authors.lastname: Piper
authors.forename: Ian
authors.initials: I
authors.lastname: Paulsen
authors.affiliation: Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology, Sydney, NSW 2007, Australia.
authors.forename: Dayong
authors.initials: D
authors.lastname: Jin
mesh_terms: D001710:Biotin / Q000737:chemistry; D003124:Colorimetry; D004247:DNA / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000378:metabolism; D007118:Immunoassay; D053758:Nanoparticles / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D001710:Biotin; D004247:DNA; D019809:Streptavidin; D006735:Horseradish Peroxidase