Table 1 Summary of ELISA’s characteristics. Main characteristics declared by the manufacturers of each ELISA kit evaluated in this study.
KIT name, reference and company | Total BDNF Quantikine ELISA #DBNT00 (R&D Systems) | Human pro-BDNF DuoSet ELISA #DY3175 (R&D Systems) | Human free BDNF Quantikine ELISA #DBD00 (R&D Systems) | Human pro-BDNF ELISA #EH4255 (FineTest) | Human BDNF ELISA #EH0043 (FineTest) | High sensitivity pro-BDNF human ELISA #SK00752-09 (Aviscera Bioscience) | High sensitivity BDNF ELISA #SK00752-01 (Aviscera Bioscience) |
|---|---|---|---|---|---|---|---|
Principle of the assay | Solid phase sandwich | Solid phase sandwich | Solid phase sandwich | Quantitative sandwich | Quantitative sandwich | Quantitative sandwich | Quantitative sandwich |
Range of detection | 15.6-1.6,000 pg/mL | 78.1-5.1,000 pg/mL | 62.5–4,000 pg/mL | 15.625-1.625,000 pg/mL | 31.25-2.25,000 pg/mL | 62.5-4.5,000 pg/mL | 3.9–250 pg/ml |
Sensitivity | 1.35 pg/mL | ND | 20 pg/mL | 9.375 pg/mL | 18.75 pg/mL | 10 pg/mL | 1 pg/mL |
Standard | Human recombinant | Human recombinant | Human recombinant | ND | ND | Human recombinant | Human recombinant |
Coating/Capture AB | Monoclonal AB specific for BDNF | Mouse anti-human pro-BDNF | Monoclonal AB specific for human free BDNF | Pre-coated AB (ND) | Pre-coated AB (ND) | Monoclonal AB against human pro-BDNF | AB against BDNF |
Primary detection AB | Monoclonal AB specific for BDNF conjugated to HRP | Biotinylated mouse anti-human pro-BDNF | Monoclonal AB specific for human free BDNF conjugated to HRP | Biotin-labelled AB (ND) | Biotinylated AB (ND) | Biotinylated monoclonal AB against human pro-BDNF | Biotinylated AB against BDNF |
Type of secondary detection | Streptavidin-HRP conjugate | Streptavidin-HRP Conjugate | Streptavidin-HRP Conjugate | Streptavidin-HRP Conjugate | Streptavidin-HRP Conjugate | ||
Recommended sample dilution | Dilution 1:100 | Optimization by the user | Dilution 1:20 | Dilution 1:100-1:200 | Dilution 1:100-1:200 | Optimization by the user | Dilution 1:160 |
Standards and samples diluent | Calibrator Diluent RD5k | PBS (#896036) + 10% FCS | Calibrator Diluent RD6P | Sample dilution buffer | Sample dilution buffer | Dilution Buffer | Dilution Buffer |
Declared species cross-reactivity | Human, mouse and rat | Human and mouse | Human | Human | Human | Human | Human and rat |
Cross-reactivity | No | 1.4% human BDNF | 13% pro-BDNF | No | No | No | < 2% pro-BDNF |
Processing time | 3–4 h | Overnight incubation + 5–6 h | 3–4 h | 4–5 h | 4 h | 6–7 h | ND |
