Table 1.
Design of experiment factors and levels investigated for DEN2
Factor | Whole plot or subplot | Level 1 | Level 2 | Level 3 | Level 4 |
---|---|---|---|---|---|
A. Plate incubation temperature | Whole plot | Ambient | 37°C | N/A | N/A |
B. Primary Ab incubation time and temperature | Whole plot | 60 minutes/37°C | Overnight/4°C | N/A | N/A |
C. Secondary Ab incubation time | Whole plot | 30 minutes | 90 minutes | N/A | N/A |
D. Cell seeding density | Subplot | 1 × 104 cells | 3 × 104 cells | N/A | N/A |
E. Multiplicity of infection | Subplot | 0.01 | 0.1 | N/A | N/A |
F. Neutralization mix incubation-time/temp | Subplot | 30 minutes/37°C | Overnight/4°C | 60 minutes/37°C | 60 minutes/ambient |
G. Primary Ab concentration | Subplot | 1 µg/mL | 10 µg/mL | N/A | N/A |
H. Secondary Ab dilution | Subplot | 1:250 | 1:1,000 | N/A | N/A |
N/A = not applicable.