Quantitative Multianalyte Microarray Immunoassay Utilizing Upconverting Phosphor Technology

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• 作者：Henna P盲kkil盲 ; Minna Ylih盲rsil盲 ; Satu Lahtinen ; Liisa Hattara ; Niina Salminen ; Riikka Arppe ; Mika Lastusaari ; Petri Saviranta ; Tero Soukka
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：October 16, 2012
• 年：2012
• 卷：84
• 期：20
• 页码：8628-8634
• 全文大小：319K
• 年卷期：v.84,no.20(October 16, 2012)
• ISSN：1520-6882

文摘

A quantitative multianalyte immunoassay utilizing luminescent upconverting single-crystal nanoparticles as reporters on an antibody array-in-well platform was demonstrated. Upconverting nanoparticles are inorganic rare earth doped materials that have the unique feature of converting low energy infrared radiation into higher energy visible light. Autofluorescence, commonly limiting the sensitivity of fluorescence-based assays, can be completely eliminated with photon upconversion technology because the phenomenon does not occur in biological materials. Biotinylated antibodies for three analytes (prostate specific antigen, thyroid stimulating hormone, and luteinizing hormone) were printed in an array format onto the bottom of streptavidin-coated microtiter wells. Analyte dilutions were added to the wells, and the analytes were detected with antibody-coated upconverting nanoparticles. Binding of the upconverting nanoparticles was imaged with an anti-Stokes photoluminescence microwell imager, and the standard curves for each analyte were quantified from the selected spot areas of the images. Single analyte and reference assays were also carried out to compare with the results of the multianalyte assay. Multiplexing did not have an effect on the assay performance. This study demonstrates the feasibility of upconverting single-crystal nanoparticles for imaging-based detection of quantitative multianalyte assays.