Table 3.

Comparison of the characteristic parameters of ADCC assays using the nanoluciferase, calcein or TR-FRET methods in the unconstrained 4-parameter logistic regression model. The EC₅₀ (means and the corresponding CV), E_min and E_max (mean and range) from the reference sample are indicated in the table for both cellular models, using the different readouts. Correlation coefficients (r²) are also indicated to illustrate the data fitting to the regression model. The relative biases (and ranges) obtained with the under- and over-concentrated test samples (80% and 125%) are also shown. The data were obtained from 3 independent experiments (see also Fig. 6A). * In the Raji/calcein model @125%, only one experiment over 3 gave data fitting with the logistic model.

		Reference sample (100%)				Under or over-concentrated test samples
Model	Method	EC50 (ng/mL) [%CV]	r2	Emin (%) [min / max]	Emax (%) [min / max]	Bias @ 80% (%) [min / max]	Bias @ 125% (%) [min / max]
Raji	Nanoluciferase	14.0 [58.4%]	0.9715	−2.7 [−5.2 / −0.6]	95.9 [93.2 / 100.0]	−5.5 [−25.1 / 7.5]	16.0 [−23.9 / 58.8]
	Calcein	53.1 [144.4%]	0.7696	−37.1 [−102.7 / 23.6]	194.1 [99.1 / 376]	532.1 [−69.1 / 1689.7]	−62.7 [NA]*
	TR-FRET	13.2 [25.9%]	0.5672	−38.16 [−57.1 / −7.4]	166.5 [80.1 / 336.6]	188.3 [−100.0 / 762.7]	−32.0 [−265.0 / 160.5]
SKOV-3	Nanoluciferase	46.6 [16.5%]	0.9753	−0.5 [−1.6 / 0.5]	52.5 [45.3 / 59.7]	−26.0 [−40.0 / −11.1]	−3.2 [−30.4 / 24.9]
	Calcein	48.51 [11.4%]	0.9113	−6.3 [−20.1 / 8.6]	93.2 [78.1 / 103.6]	50.5 [6.9 / 123.5]	14.8 [−5.2 / 38.3]
	TR-FRET	25.5 [12.1%]	0.9684	0.33 [−2.8 / 4.1]	90.6 [86.6 / 94.1]	−45.2 [−50.5 / −35.2]	19.3 [1.0 / 38.0]