sFIDA automation yields sub-femtomolar limit of detection for Aβ aggregates in body fluids

Herrmann, Yvonne; Kulawik, Andreas; Kühbach, Katja; Hülsemann, Maren; Peters, Luriano; Bujnicki, Tuyen; Kravchenko, Kateryna; Linnartz, Christina; Willbold, Johannes; Zafiu, Christian; Bannach, Oliver; Willbold, Dieter

doi:10.1016/j.clinbiochem.2016.11.001

Cited by 14 publications

(15 citation statements)

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“…To develop a suitable method to investigate target engagement in vivo, an assay able to quantitate Aβ oligomers in body liquids (e.g., cerebrospinal fluid, plasma) was developed. The assay, called sFIDA, is insensitive to Aβ monomers and achieves single-particle sensitivity [26, 29, 30]. In the present study, we developed this technique further in order to measure Aβ oligomers in organ homogenates.…”

Section: Resultsmentioning

confidence: 99%

“…To develop an experiment that allows the investigation of target engagement in vivo, we decided to use brain homogenates without enrichment steps for human Aβ, as this could potentially lead to the destruction of native Aβ oligomers and also to the formation of artificial Aβ aggregates formed during Aβ precipitation steps. Because the brain homogenates contain not only Aβ, but also all other brain-derived components, we used the ultra-sensitive and specific sFIDA assay [26, 29–31] for the quantification of Aβ oligomers in the DGC-fractionated brain homogenates ex vivo. The most significant reduction in Aβ containing particles by RD2 treatment was observed in fraction 10, which corresponds to particles with a molecular weight of larger than 400 kDa [14].…”

Section: Discussionmentioning

confidence: 99%

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Aβ Oligomer Elimination Restores Cognition in Transgenic Alzheimer’s Mice with Full-blown Pathology

et al. 2018

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Oligomers of the amyloid-β (Aβ) protein are suspected to be responsible for the development and progression of Alzheimer's disease. Thus, the development of compounds that are able to eliminate already formed toxic Aβ oligomers is very desirable. Here, we describe the in vivo efficacy of the compound RD2, which was developed to directly and specifically eliminate toxic Aβ oligomers. In a truly therapeutic, rather than a preventive study, oral treatment with RD2 was able to reverse cognitive deficits and significantly reduce Aβ pathology in old-aged transgenic Alzheimer's Disease mice with full-blown pathology and behavioral deficits. For the first time, we demonstrate the in vivo target engagement of RD2 by showing a significant reduction of Aβ oligomers in the brains of RD2-treated mice compared to placebo-treated mice. The correlation of Aβ elimination in vivo and the reversal of cognitive deficits in old-aged transgenic mice support the hypothesis that Aβ oligomers are relevant not only for disease development and progression, but also offer a promising target for the causal treatment of Alzheimer's disease.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Aβ Oligomer Elimination Restores Cognition in Transgenic Alzheimer’s Mice with Full-blown Pathology

et al. 2018

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“…As a calibration standard bioconjugated silica nanoparticles provide interassay comparability and translation from pixel count to molar Aβ oligomer concentration. The automation of the sFIDA assay results in improved intra‐assay precision, linearity and sensitivity in comparison to the manual application, and achieves a limit of detection in the subfemtomolar range .…”

Section: Discussionmentioning

confidence: 99%

“…Since then, the sFIDA method underwent fundamental optimization in respect to automation , adaptation to blood‐based analysis and development of a suitable calibration standards .…”

Section: Main Textmentioning

confidence: 99%

Advancements of the sFIDA method for oligomer‐based diagnostics of neurodegenerative diseases

et al. 2018

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Early diagnosis of Alzheimer's disease (AD) is of great importance for the development of therapeutics and their application in the clinical environment. Amyloid β (Aβ) oligomers are crucial for the onset and progression of AD and represent a popular drug target, being presumably the most direct biomarker. Efforts to measure Aβ oligomers in body fluids are hampered by the low analyte concentration and presence of Aβ monomers. The surface‐based fluorescence intensity distribution analysis (sFIDA) features both highly specific and sensitive oligomer quantitation as well as total insensitivity towards monomers. In this Review, we highlight structural features of oligomeric and fibrillar Aβ. Recent advancements in sFIDA assay development have been the successful automation, adaption for additional biomarkers such as α‐synuclein oligomers, and significant improvement of essential assay parameters.

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“…Moreover, monomers are present in great excess, necessitating that assays have a high selectivity for oligomers. To overcome these challenges in detecting aggregated α-synuclein assemblies, we used the sFIDA (surface-based fluorescence intensity distribution analysis) assay, which uses antibodies targeting overlapping or identical epitopes, enabling it to specifically detect aggregates in the presence of monomers [40][41][42][43] . Here, the Syn211 antibody that recognizes amino acids 121-125 on α-synuclein is attached to a glass surface and captures α-synuclein species in stool homogenates 44 .…”

Section: Introductionmentioning

confidence: 99%

Patients with isolated REM-sleep behavior disorder have elevated levels of alpha-synuclein aggregates in stool

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et al. 2023

npj Parkinsons Dis.

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Misfolded and aggregated α-synuclein is a neuropathological hallmark of Parkinson’s disease (PD). Thus, α-synuclein aggregates are regarded as a biomarker for the development of diagnostic assays. Quantification of α-synuclein aggregates in body fluids is challenging, and requires highly sensitive and specific assays. Recent studies suggest that α-synuclein aggregates may be shed into stool. We used surface-based fluorescence intensity distribution analysis (sFIDA) to detect and quantify single particles of α-synuclein aggregates in stool of 94 PD patients, 72 isolated rapid eye movement sleep behavior disorder (iRBD) patients, and 51 healthy controls. We measured significantly elevated concentrations of α-synuclein aggregates in stool of iRBD patients versus those of controls (p = 0.024) or PD patients (p < 0.001). Our results show that α-synuclein aggregates are excreted in stool and can be measured using the sFIDA assay, which could support the diagnosis of prodromal synucleinopathies.

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sFIDA automation yields sub-femtomolar limit of detection for Aβ aggregates in body fluids

Cited by 14 publications

References 14 publications

Aβ Oligomer Elimination Restores Cognition in Transgenic Alzheimer’s Mice with Full-blown Pathology

Aβ Oligomer Elimination Restores Cognition in Transgenic Alzheimer’s Mice with Full-blown Pathology

Advancements of the sFIDA method for oligomer‐based diagnostics of neurodegenerative diseases

Patients with isolated REM-sleep behavior disorder have elevated levels of alpha-synuclein aggregates in stool

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