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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
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Alzheimer's Research & Therapy

ISSN: 1758-9193