Sample Nominal Conc. (ng/mL) 992 mAb Calculated Conc. (ng/mL) 992 mAb %BIAS 1024 mAb Calculated Conc. (ng/mL) 1024 mAb %BIAS 1024 mAb at ULOQ 992 mAb at ULOQ Sum of 992 mAb and 1024 mAb at ULOQ Sum of 1024 mAb and 992 mAb at ULOQ Total calculated conc. (ng/mL) %BIAS
  Std 1 150.0 158.0 105.3 ­- ­- 11051.6 ­- 11209.6 ­- 11120.5 99.2
Set 1 Std 2 250.0 263.0 105.2 ­- ­- 10426.9 ­- 10689.9 ­- 11502.6 107.6
  Std 3 500.0 590.0 118.0 ­- ­- 10491.9 ­- 11081.9 ­- 12180.1 109.9
  Std 4 1000.0 1146.6 114.7 ­- ­- 10154.0 ­- 11300.6 ­- 12687.6 112.3
  Std 1 150.0 ­- ­- 161.6 107.8 ­- 11730.0 ­- 11891.6 12320.1 105.0
Set 2 Std 2 250.0 ­- ­- 239.0 95.6 ­- 13011.3 ­- 13250.3 12249.6 94.1
  Std 3 500.0 ­- ­- 434.3 86.9 ­- 14181.3 ­- 14615.6 9934.21 70.1
  Std 4 1000.0 ­- ­- 997.4 99.7 ­- 10596.9 ­- 11594.3 12312.7 116.2
Sample 992 and 1024 mAb nominal conc. (ng/mL) 1024 mAb calculated conc. (ng/mL) 992 mAb calculated conc. (ng/mL) Sum of 1024 mAb and 992 mAb calculated conc. (ng/mL) Total mAbs calculated conc. (ng/mL) %Bias
VS-LLOQ_1 150.0 165.8 141.5 307.2 301.6 98.2
VS-LLOQ_2 150.0 167.1 144.6 311.7 288.6 92.6
VS-LLOQ_3 150.0 173.1 162.1 335.3 290.1 86.5
VS-LLOQ_4 150.0 164.6 117.2 281.8 288.8 102.5
VS-LLOQ_5 150.0 152.4 152.2 304.6 297.7 97.7
VS-LLOQ_6 150.0 125.5 160.0 285.5 310.9 108.9
VS-LLOQ_7 150.0 141.8 148.9 290.7 263.1 90.5
VS-LLOQ_8 150.0 162.0 153.6 315.6 264.0 83.6
VS-LLOQ_9 150.0 162.5 156.7 319.2 313.8 98.3
VS-LLOQ_10 150.0 150.7 164.1 314.7 266.1 84.5
VS-LLOQ_11 150.0 152.6 158.2 310.7 267.0 85.9
VS-Low_1 450.0 498.5 460.1 958.5 942.4 98.3
VS-Low_2 450.0 509.0 490.1 999.1 1074.1 107.5
VS-Low_3 450.0 479.4 469.8 949.2 1016.4 107.1
VS-Low_4 450.0 530.8 477.9 1008.7 956.2 94.8
VS-Low_5 450.0 511.5 513.8 1025.3 1120.3 109.3
VS-Low_6 450.0 447.1 479.5 926.7 1007.3 108.7
VS-Low_7 450.0 414.8 450.2 865.0 783.9 90.6
VS-Low_8 450.0 462.6 458.4 921.0 873.7 94.9
VS-Low_9 450.0 393.5 424.1 817.6 1019.4 124.7
VS-Low_10 450.0 446.2 523.4 969.6 1124.5 116.0
VS-Low_11 450.0 461.5 377.6 839.1 1072.3 127.8
VS-Medium_1 1500.0 DEV DEV DEV DEV DEV
VS-Medium_2 1500.0 1701.1 1837.1 3538.3 3181.9 89.9
VS-Medium_3 1500.0 1634.9 1750.2 3385.1 3569.9 105.5
VS-Medium_4 1500.0 1717.1 1782.8 3499.9 3593.0 102.7
VS-Medium_5 1500.0 1706.7 1614.5 3321.1 3235.2 97.4
VS-Medium_6 1500.0 1475.4 1632.8 3108.2 2586.1 83.2
VS-Medium_7 1500.0 1524.1 1389.1 2913.1 2789.6 95.8
VS-Medium_8 1500.0 1443.1 1424.3 2867.4 2832.3 98.8
VS-Medium_9 1500.0 1697.3 1497.9 3195.1 3387.2 106.0
VS-Medium_10 1500.0 1698.5 1722.1 3420.6 3285.3 96.0
VS-Medium_11 1500.0 1875.6 1792.7 3668.3 3591.1 97.9
VS-High_1 8500.0 8401.0 8187.5 16588.5 15863.2 95.6
VS-High_2 8500.0 8948.2 9003.0 17951.2 16840.3 93.8
Sample 992 and 1024 mAb nominal conc. (ng/mL) 1024 mAb calculated conc. (ng/mL) 992 mAb calculated conc. (ng/mL) Sum of 1024 mAb and 992 mAb calculated conc. (ng/mL) Total mAbs calculated conc. (ng/mL) %Bias
VS-High_3 8500.0 8501.8 8986.3 17488.1 17506.2 100.1
VS-High_4 8500.0 8797.9 8114.8 16912.7 19210.4 113.6
VS-High_5 8500.0 8881.8 8691.8 17573.6 18023.8 102.6
VS-High_6 8500.0 8191.7 8791.02 16982.8 14244.0 83.9
VS-High_7 8500.0 8800.2 8909.84 17710.0 15840.6 89.4
VS-High_8 8500.0 8421.5 9575.83 17997.4 14445.5 80.3
VS-High_9 8500.0 8429.3 8714.6 17143.8 17328.8 101.1
VS-High_10 8500.0 8183.6 9253.3 17436.9 16197.1 92.9
VS-High_11 8500.0 8406.3 8774.3 17180.6 16795.2 97.8
VS-ULOQ_1 10000.0 11091.2 13600.9 24692.1 22622.1 91.6
VS-ULOQ_2 10000.0 11414.5 11984.6 23399.1 24885.9 106.4
VS-ULOQ_3 10000.0 10760.8 11311.2 22072.0 21742.9 98.5
VS-ULOQ_4 10000.0 11190.5 12435.0 23625.5 22692.7 96.1
VS-ULOQ_5 10000.0 11018.2 14001.5 25019.7 24498.1 97.9
VS-ULOQ_6 10000.0 10099.1 11867.0 21966.1 16253.0 74.0
VS-ULOQ_7 10000.0 11463.1 11306.4 22769.5 20383.4 89.5
VS-ULOQ_8 10000.0 11299.2 10712.1 22011.3 17992.2 81.7
VS-ULOQ_9 10000.0 11412.3 11301.9 22714.2 23017.8 101.3
VS-ULOQ_10 10000.0 11870.9 11608.8 23479.7 24067.1 102.5
VS-ULOQ_11 10000.0 10427.4 11056.1 21483.5 21557.2 100.3
Table 8: Evaluation of total mAbs accuracies when compared to the sum of individual mAbs measured concentrations. DEV stands for deviation from the sample preparation procedure. In Set1 and Set2 are also reported the specificity experiment results where one mAb at ULOQ is spiked over a concentration curve of the other mAb. The last column reports the accuracy of the calculated total mAb concentrations (%BIAS) where the nominal concentration was assumed to be the sum of the individual mAbs calculated concentrations. Out of accuracy acceptance criteria, i.e. ±20% BIAS% (±25%BIAS % at LLOQ) are shown in bold.