Abstract
Brain-Derived Neurotrophic Factor (BDNF) has attracted increasing interest as potential biomarker to support the diagnosis or monitor the efficacy of therapies in brain disorders. Circulating BDNF can be measured in serum, plasma or whole blood. However, the use of BDNF as biomarker is limited by the poor reproducibility of results, likely due to the variety of methods used for sample collection and BDNF analysis. To overcome these limitations, using sera from 40 healthy adults, we compared the performance of five ELISA kits (Aviscera-Bioscience, Biosensis, Millipore-ChemiKineTM, Promega-Emax®, R&D-System-Quantikine®) and one multiplexing assay (Millipore-Milliplex®). All kits showed 100% sample recovery and comparable range. However, they exhibited very different inter-assay variations from 5% to 20%. Inter-assay variations were higher than those declared by the manufacturers with only one exception which also had the best overall performance. Dot-blot analysis revealed that two kits selectively recognize mature BDNF, while the others reacted with both pro-BDNF and mature BDNF. In conclusion, we identified two assays to obtain reliable measurements of human serum BDNF, suitable for future clinical applications.
Highlights
In the present study we propose a highly reproducible Brain-Derived Neurotrophic Factor (BDNF) quantification method potentially suitable for clinical applications
To obtain a reduction in the variability of measurements due to technical issues, we identified the best performing assay for BDNF detection by comparing six widely used commercial kits based on the ELISA technique, the most common way to measure circulating BDNF and, according to our results, one of the main sources of inconsistencies among studies
The BDNF quantification from plasma can be sensitive to preparation procedures and is very difficult to be reproducible among different operators
Summary
Recent exploratory studies have found increased serum BDNF levels following holistic neuro-rehabilitative approaches, including computer-assisted cognitive enhancement in schizophrenia[20], aerobic exercise in stroke[21] and mindfulness clinical trials in bipolar-disorder[22]. These studies showed differences in collecting the samples that may add an additional variability other than the well known variables, such as BMI, drugs, smoking, etc.[23], affecting circulating BDNF levels, resulting in further difficulties in assessing BDNF levels related to the pathology or treatment. We measured BDNF concentration in sera from 40 healthy volunteers using 6 different commercial kits and comparing their performances
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