Please Wait...

Analytical Performance Characteristics of a Novel Immunoassay for the Quantification of BACE-1 in Human Cerebrospinal Fluid

Emeric Chassaing, BS1, Mélanie Bodnar-Wachtel, PhD1, Tanja Schubert, PhD1, Hugo Marcel Vanderstichele, PhD2, Erik Stoops, Eng2 and Philippe Vergnaud, MSc1, (1)Bioclinica Lab, Lyon, France, (2)ADx NeuroSciences, Gent, Belgium

 

BACKGROUND

Accumulation of amyloid-β (Aβ) into plaques in the brain is one of the hallmarks of Alzheimer’s Disease (AD). This neurotoxic peptide is produced by the consecutive cleavage of the amyloid precursor protein (APP) by β-site APP cleaving enzyme 1 (BACE1) and γ-secretase. Due to its key role in amyloid plaque formation, BACE-1 is a major therapeutic target for reducing the levels of amyloid in AD brain. The BACE1 ELISA quantifies specifically BACE-1, in contrast to BACE-1 enzyme activity assays, which suffer from a BACE-2 interference. This study documents the analytical performance characteristics of a novel ELISA, targeting BACE-1 protein in human Cerebrospinal Fluid (CSF).

METHODS

The BACE-1 ELISA was developed with two monoclonal antibodies. Lyophilized calibrators, as well as run-validation control samples were included in the final kit format. Total assay times is 4 hours. After having performed familiarization runs and adaptation of the test procedures to the needs of a service provider, the assay was challenged internally for its analytical performance characteristics (e.g., precision, parallelism, spike-recovery, working range, sample stability) by using commercially available CSF samples (n=34).

RESULTS

The assay was qualified for measurement of BACE-1 protein in undiluted CSF and EDTA-plasma. Full validation was done for the CSF assay format. The new colorimetric ELISA specifically quantifies BACE-1 in the absence of matrix interference (parallelism 93 % ± 10 %; dilution range: neat – 1/16). Spike/recovery testing revealed good recovery rates (103 % ± 6 %). The precision resulted in intra- and inter-assay variability (% CV) below 5 % and 11 %, respectively. The working range is confirmed between 272 pg/mL (Lower Limit of Quantitation) to 10,752 pg/mL (Upper Limit of Quantitation) and lowest concentration detectable (LOD) is 29.2 pg/mL. CSF stability at +4°C and room temperature is acceptable after 24 hours of storage; consecutive freezing/thawing of CSF up to 4 cycles did not affect BACE-1 concentrations.

CONCLUSIONS

The newly developed BACE-1 colorimetric ELISA assay meets all internal acceptance criteria for clinical trial sample testing and represents an additional tool for patient management.

LEARN MORE OR SPEAK WITH OUR EXPERTS

CONTACT US
Leader in Clinical Trial
Management Solutions

Successful clinical trials require the ability to see key details and uncover hidden insights. Bioclinica utilizes science and technology to bring clarity to clinical trials, helping companies to develop new life-improving therapies more efficiently and safely.

Learn more about the role of #CROs in Payment Management, desired characteristics of Payment System and Technology-… https://t.co/gsEvSwwHTR
Bioclinica (2 weeks ago)
Bioclinica is an agile organization with more 2,300 colleagues around the globe - and growing! Learn more here.… https://t.co/IfyE3B7fba
Bioclinica (2 weeks ago)
Immediate opening for a Citrix Administrator in Princeton, NJ . Must have previous experience with citrix and windo… https://t.co/YYw9Y4o6m0
Bioclinica (2 weeks ago)
Going to DIA Japan in Toyko? Meet our team at booth 47! https://t.co/3Sdi4Q2dIs https://t.co/GVecVewIgP
Bioclinica (3 weeks ago)
Meet Bioclinica’s expert Jeffrey Heilbraun, M.S. to learn more about Cardiac Safety Services. #bioclinica… https://t.co/iBAv471ZGK
Bioclinica (3 weeks ago)
Immediate opening for a Clinical Project Manager in Princeton, NJ – Project Management experience within a CRO is h… https://t.co/mVtp6nayS0
Bioclinica (4 weeks ago)