ProteinSimple eBook - 28

Simplifying Protein Analysis * Multi-Analyte, Automated, Microﬂuidic Immunoassay Platform
A
A

BB

Sample

IL-1β Calibration Curve

Signal (RFU/ms)

1,000

Running
Buffer

100

10

GNRs
1.0

Waste

0

1

CC

10

100

Concentration, pg/mL

1,000

10,000

1,000

10,000

1,000

10,000

IL-6 Calibration Curve

10,000

Detect Fluor

IL-1β

IL-6

IL-10 Variable

PANEL 1

PANEL 2

PANEL 3

PANEL 4

Channel 1

IL-1β

IL-1β

IL-1β

IL-1β

Channel 2

IL-6

IL-6

IL-6

IL-6

Channel 3

IL-10

IL-10

IL-10

TNF-α

IL-2

IL-5

IL-12

1.0

0

1

D
D

10

100

Concentration, pg/mL

IL-10 Calibration Curve

10,000

100%

1,000

80%
60%
40%

100
10
1.0

20%
0%
IL-1β

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10

Signal (RFU/ms)

Recovery of 40 pg/mL

E E 120%

28

100

IL-10

Channel 4

Signal (RFU/ms)

1,000

IL-6

IL-10

TNF-α

IL-2

IL-5

IL-12

0.1
0

1

10

100

Concentration, pg/mL

Figure 4. Assay uniformity as influenced by panel
composition. (A) Schematic of circuit composition across the
four panels. (B-D) Calibration curve stability for each of the
constant assays. Each graph contains the mean standard
curve with standard deviation error bars; the maximum CV
between standard curve points was 4%,7%,and 7% for
IL-1β, IL-6, and IL-10, respectively. (E) Recovery of a
40 pg/mL calibration point; the results were averaged
across the four panels lL-l, lL-6, and IL-10; and single
measurements for TNF-α, lL-2, lL-5, and lL-12.
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