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000823896 1001_ $$0P:(DE-Juel1)159122$$aHerrmann, Yvonne$$b0$$ufzj
000823896 245__ $$asFIDA automation yields sub-femtomolar limit of detection for Aβ aggregates in body fluids
000823896 260__ $$aAmsterdam [u.a.]$$bElsevier Science$$c2017
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000823896 520__ $$aObjectives: Alzheimer's disease (AD) is a neurodegenerative disorder with yet non-existent therapeutic and limited diagnostic options. Reliable biomarker-based AD diagnostics are of utmost importance for the development and application of therapeutic substances. Wehave previously introduced a platformtechnology designated ‘sFIDA’ for the quantitation of amyloid β peptide (Aβ) aggregates as AD biomarker. In this study we implemented the sFIDA assay on an automated platform to enhance robustness and performance of the assay.Design and methods: In sFIDA (surface-based fluorescence intensity distribution analysis) Aβ species are immobilized by a capture antibody to a glass surface. Aβ aggregates are then multiply loaded with fluorescent antibodies and quantitated by high resolution fluorescence microscopy. As a model system for Aβ aggregates, weused Aβ-conjugated silica nanoparticles (Aβ-SiNaPs) diluted in PBS buffer and cerebrospinal fluid, respectively. Automation of the assay was realized on a liquid handling system in combination with a microplate washer.Results: The automation of the sFIDA assay results in improved intra-assay precision, linearity and sensitivity in comparison to the manual application, and achieved a limit of detection in the sub-femtomolar range.Conclusions: Automation improves the precision and sensitivity of the sFIDA assay, which is a prerequisite for high-throughput measurements and future application of the technology in routine AD diagnostics.
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000823896 7001_ $$0P:(DE-Juel1)162310$$aKulawik, Andreas$$b1$$ufzj
000823896 7001_ $$0P:(DE-Juel1)145440$$aKühbach, Katja$$b2$$ufzj
000823896 7001_ $$0P:(DE-Juel1)145968$$aHülsemann, Maren$$b3
000823896 7001_ $$0P:(DE-Juel1)145123$$aPeters, Luriano$$b4$$ufzj
000823896 7001_ $$0P:(DE-Juel1)162212$$aBujnicki, Tuyen$$b5$$ufzj
000823896 7001_ $$0P:(DE-Juel1)136806$$aKravchenko, Kateryna$$b6
000823896 7001_ $$0P:(DE-Juel1)145159$$aLinnartz, Christina$$b7$$ufzj
000823896 7001_ $$0P:(DE-Juel1)159192$$aWillbold, Johannes$$b8$$ufzj
000823896 7001_ $$0P:(DE-Juel1)162137$$aZafiu, Christian$$b9$$ufzj
000823896 7001_ $$0P:(DE-Juel1)157832$$aBannach, Oliver$$b10$$ufzj
000823896 7001_ $$0P:(DE-Juel1)132029$$aWillbold, Dieter$$b11$$eCorresponding author
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