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Table 14 Assay characteristics compared to existing commercial ELISA assays for mice

From: Development of a novel, sensitive translational immunoassay to detect plasma glial fibrillary acidic protein (GFAP) after murine traumatic brain injury

Supplier MyBioSource Abcam* ELISA
Genie
Millipore Cusabio Novel MSD assay
Catalog number MBS723651 MBS2515511 MBS2018967 MBS763852 MBS2701012 ab233621 MOFI00191 NS830 E08603m  
Cost/plate $640 USD $490 USD $625 USD $415 USD $350 USD $593 CAD €599 $693 CAD $600 USD $150 USD
Volume of sample (μL) 50 100 100 100 100 50 100 100 50–100 12.5
Detection range (pg/mL) 50–1000 15.63–1000 15.6–1000 15.6–1000 62.5–4000 125–8000 15.6–1000 1500–100,000 3.12–200 24.8–16,533.9
LLOD (pg/mL) Outcome 1 9.38 < 6.3 9.38 23.3 8.7 9.375 1500 0.78 9
Protocol N/A N/A 2 SD above background of 20 blanks replicates. N/A 2 SD above background of 20 blank replicates. N/A N/A N/A   2.5 SD above background 16 blank replicates
Intra-assay CV (%) Outcome < 9 4.6 < 10 < 8 < 10 4.4 < 8 6.8 < 8 5.0
Protocol N/A Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-run CV from 1 run, 8 replicates. Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-run CV from 1 run, 3 specimens, 16 replicates/specimen. Within-run CV from 1 run, 3 specimens, 20 replicates/specimen. Within-day CV from 5 days, 1 run/day, 3 plasma pools/run, 5 replicates/pool.
Inter-assay CV (%) Outcome < 10 5.1 < 12 < 10 < 12 5.8 < 10 5.2 < 10 7.2
Protocol N/A Between-run CV from 3 runs, 3 specimens/run, 20 replicates/specimen. Between-run CV from 3 runs, 3 specimens/run, 8 replicates/specimen. Between-run CV from 3 runs, 3 specimens/run, 8 replicates/specimen. Between-run CV from 3 runs, 3 specimens/run, 8 replicates/specimen. Between-run CV from 3 runs. Between-run CV from 3 runs, 3 specimens/run, 8 replicates/specimen. Between-run CV from 3 runs, 6 specimens/run, 2 replicates/specimen. Between-run CV from 20 runs, 3 specimens/run. Between-day CV from 5 runs, 3 plasma pools/run, 5 replicates/pool.
Recovery Outcome 94–103% 89–103% 78–99% 85–105% 80–98% 95–100% 88–105% 95–109% 90–104% 62–97%A
86–105%B
Protocol N/A Spiked serum, plasma, culture media (n = 5 each) at 3 concentrations. Spiked plasma (n = 10) and serum (n = 5). Spiked plasma (n = 10) and serum (n = 5). Spiked plasma (n = 10) and serum (n = 5). Spiked brain extract. Spiked plasma (n = 10) and serum (n = 5). Spike brain (n = 1) or cell (n = 2) lysate with 1.56–50 ng GFAP. Spiked serum (n = 5) or plasma (n = 5). Spike plasma (n = 5) with 100, 1000, and 10,000 pg/mL GFAP.
Dilution linearity Outcome 92–108% 80–109% 80–108% 81–104% 78–105% 87–100% 81–104% 77–122% 85–108% 93–105%
Protocol 2× to 16× Spiked serum, plasma, culture media (n = 5 each), diluted 1:2 to 1:16. Spiked plasma (n = 10) and serum (n = 5), diluted 1:2 to 1:16. Spiked plasma (n = 10) and serum (n = 5), diluted 1:2 to 1:16. Spiked plasma (n = 10) and serum (n = 5), diluted 1:2 to 1:16. Brain extract (n = 2) diluted 1:2 to 1:16. Spiked plasma (n = 10) and serum (n = 5), diluted 1:2 to 1:16. Spiked brain lysate (n = 3) 1:3 to 1:27. Spiked serum (n = 4), diluted 1:1 to 1:8. Plasma (n = 3) diluted 1:2 to 1:16.
  1. *Same antibody pair used between assays. A Recovery data for samples with low endogenous GFAP. B Recovery data for samples with high endogenous GFAP. LLOD lower limit of detection, CV coefficient of variation, SD standard deviation. Data retrieved from supplier product websites