Table 2.

Assay characteristics, methods, and summarized observations. Assay characteristics are reported along with methods and details for the five HIV bnAbs tested.

	Experimental design		10–1074	3BNC117	3BNC117LS	PGDM1400	PGT121
Cut Point	Treatment-naïve HIV+ and HIV- subjects' serum samples were assayed	Fixed	2.7 S/B	1.7 S/B	2.2 S/B	1.45 S/B	3.0 S/B
		Floating	+0.7 S/B	+0.4 S/B	+0.3 S/B	+0 S/B	+0.7 S/B
Specificity	Serum samples from three different categories of patients (HIV-, HIV+ ART+, and HIV+ ART-) were tested in duplicates, and then retested with additional anti-id spiked-in.		All samples became ADA positive upon anti-id spike in	All samples became ADA positive upon anti-id spike in	All samples became ADA positive upon anti-id spike in	All samples became ADA positive upon anti-id spike in	HIV+ ART+ and HIV- samples all became ADA positive ART- subjects had reduced anti-id induced signal. (5/16 remained below cut point)
	Serum samples were spiked with 3200 ng/mL of an unrelated mAb or 20 IU/mL of rheumatoid factor (RF) and assayed.		2 of 13 samples became positive upon addition of either mAb or RF.	0 of 13 samples became positive.	0 of 12 samples became positive.	0 of 10 samples became positive.	6 of 13 samples became positive upon addition of anti-PGDM1400 All 13 samples became positive upon addition of RF
Drug tolerance	PNHS containing anti-id at concentrations sufficient to result in a positive response call were spiked with the respective mAb over a range of concentrations and assayed in duplicate by two operators. An unrelated mAb was also spiked into samples containing the mAb anti-id.	100 ng/mL anti-ida	70 μg/mL	10 μg/mL	30 μg/mL	12.5 μg/mL	1.9 μg/mL
		500 ng/mL anti-id	>140 μg/mL	>60 μg/mL	>60 μg/mL	>40 μg/mL	1.3 μg/mL
		Unrelated mAb	140 μg/mL 3BNC117 did not result in any signal interference	80 μg/mL 10–1074 did not result in any signal interference	60 μg/mL 10–1074 did not result in any signal interference	40 μg/mL PGT121 did not result in any signal interference	20 μg/mL PGDM1400 did not result in any signal interference
Stability	QC samples were subjected to either freeze-thaw cycles, three hours at room temperature, or three days at 4 °C, then compared to samples that were stored at −80 °C by two operators	Stability of the samples was tested through three freeze-thaw cycles, for three hours at room temperature (RT), and for three days at 4 °C.	RT: 3 h 4 °C: 3 days −80 °C: 3 months	RT: 3 h 4 °C: 3 days −80 °C: 3 months	RT: 3 h 4 °C: 3 days −80 °C: 3 months	RT: 3 h 4 °C: 3 days −80 °C: 3 months	RT: 3 h 4 °C: 3 days −80 °C: 7 months

^aPGT 121 drug tolerance was established with a spike in of 200 ng/mL anti-Id.