TY - JOUR
AU - Herrmann, Yvonne
AU - Kulawik, Andreas
AU - Kühbach, Katja
AU - Hülsemann, Maren
AU - Peters, Luriano
AU - Bujnicki, Tuyen
AU - Kravchenko, Kateryna
AU - Linnartz, Christina
AU - Willbold, Johannes
AU - Zafiu, Christian
AU - Bannach, Oliver
AU - Willbold, Dieter
TI - sFIDA automation yields sub-femtomolar limit of detection for Aβ aggregates in body fluids
JO - Clinical biochemistry
VL - 50
IS - 4-5
SN - 0009-9120
CY - Amsterdam [u.a.]
PB - Elsevier Science
M1 - FZJ-2016-06528
SP - 244-247
PY - 2017
AB - Objectives: 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.
LB - PUB:(DE-HGF)16
UR - <Go to ISI:>//WOS:000395965100015
C6 - pmid:27823959
DO - DOI:10.1016/j.clinbiochem.2016.11.001
UR - https://juser.fz-juelich.de/record/823896
ER -