Abstract
Introduction: Brain-Derived Neurotrophic Factor (BDNF) is a neurotrophin that plays an essential role in neuronal development and synaptic plasticity. Enzyme-linked immunosorbent assay (ELISA) kits can be used to measure BDNF in blood serum. However, information about sample processing and appropriate standards when clotting factors are present in the sample remains scattered. We aim at validating and optimizing the BDNF ELISA kit with respect to measurements in heparin-plasma blood samples.
Methods: blood heparin-treated plasma from 28 adults were processed as the commercial kit Mature BDNF/proBDNF Combo Rapid ELISA Kit (BEK-2211/2237, Biosensis, Australia) suggested for human plasma.
Results: we found that the pre-analytical conditions were critical for plasma samples. Determination of the intra assay variation and the accuracy and yield of the BDNF ELISA kit in heparin-plasma samples were conducted with the optimal dilutions. A linear dilution curve determined the optimal dilutions of plasma extracts; we found that 1:2 dilution optimized the detection levels for proBDNF and 1:50 dilution for matureBDNF. The range of sensitivity for BDNF isoforms in heparin-plasma samples was accurate to the manufacturer’s descriptions. Furthermore, we established the coefficient of variation (CV%) between pro- and mature BDNF. The inter-assay variation showed a CV% of 5% for mature BDNF and 25% for proBDNF.
We conclude that the BDNF ELISA kit determines heparin-plasma BDNF accurately and with high reproducibility. Last, it can be used for the measurement of BDNF isoforms in samples with clotting agents. Comparisons between clotting-plasma and serum samples will be determined and presented as part of this pilot study.
Methods: blood heparin-treated plasma from 28 adults were processed as the commercial kit Mature BDNF/proBDNF Combo Rapid ELISA Kit (BEK-2211/2237, Biosensis, Australia) suggested for human plasma.
Results: we found that the pre-analytical conditions were critical for plasma samples. Determination of the intra assay variation and the accuracy and yield of the BDNF ELISA kit in heparin-plasma samples were conducted with the optimal dilutions. A linear dilution curve determined the optimal dilutions of plasma extracts; we found that 1:2 dilution optimized the detection levels for proBDNF and 1:50 dilution for matureBDNF. The range of sensitivity for BDNF isoforms in heparin-plasma samples was accurate to the manufacturer’s descriptions. Furthermore, we established the coefficient of variation (CV%) between pro- and mature BDNF. The inter-assay variation showed a CV% of 5% for mature BDNF and 25% for proBDNF.
We conclude that the BDNF ELISA kit determines heparin-plasma BDNF accurately and with high reproducibility. Last, it can be used for the measurement of BDNF isoforms in samples with clotting agents. Comparisons between clotting-plasma and serum samples will be determined and presented as part of this pilot study.
| Original language | American English |
|---|---|
| Pages | p. 18 |
| State | Published - 22 Feb 2021 |
| Event | Oklahoma State University Center for Health Sciences Research Days 2021: Poster presentation - Oklahoma State University Center for Health Sciences Campus, Tulsa, United States Duration: 22 Feb 2021 → 26 Feb 2021 |
Conference
| Conference | Oklahoma State University Center for Health Sciences Research Days 2021 |
|---|---|
| Country/Territory | United States |
| City | Tulsa |
| Period | 22/02/21 → 26/02/21 |
Keywords
- Brain-Derived Neurotrophic Factor
- BDNF
- Enzyme-linked immunosorbent assay
- ELISA kit
- blood serum