The performance of photon-upconversion nanoparticles (UCNPs) as background-free luminescent labels in bioanalytical applications strongly depends on the preparation of well-defined and water-dispersible nanoconjugates. We have exploited the separation power of agarose-gel electrophoresis to purify milligram amounts of homogeneous UCNPs covered with carboxylated silica, biotin, or streptavidin with recovery rates of 30 to 50%. Clusters containing discrete numbers of UCNPs were isolated from the gel and reanalyzed by agarose-gel electrophoresis, single-nanoparticle-upconversion microscopy, and additional complementary methods. The purified nanoconjugates improved conventional (analogue) bioaffinity assays and provided highly monodisperse conjugates for assays that rely on counting individual UCNPs (digital assays).

Centre for Modern Interdisciplinary Technologies Chair of Environmental Chemistry and Bioanalytics Faculty of Chemistry Nicolaus Copernicus University in Toruń 87 100 Toruń Poland

Department of Biochemistry Biotechnology University of Turku 20014 Turku Finland

Institute of Analytical Chemistry Chemo and Biosensors University of Regensburg 93053 Regensburg Germany

Institute of Analytical Chemistry of the Czech Academy of Sciences 602 00 Brno Czech Republic

Institute of Physics of Materials of the Czech Academy of Sciences 616 62 Brno Czech Republic

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