Table 3.

Exploratory panning of ALTHEA Gold libraries™ with three disclosed targets.

			Unique clonesc				KD (nM)f
Target	Selectiona	Hit rateb	SL1	SL2	Yield (mg/L) d	Monomeric Content (%)e	Fab	IgG	Tm (°C)g
TNFα	IM, R4, EA	20%	1	1	50–60	92–97	590	0.094–8	77–80
HSA	EP, R3, AE	83%	-	3	90	95	3.3	0.8	81
HEL	MB, R3, AE	6%-68%	3	30	-	-	-	-	-

^aIM: Immunotubes; R4: Four rounds of panning; EA: Elution with Adalimumab; EP: ELISA plates; R3: Three rounds of panning; AE: Acid Elution; MB: Magnetic beads.

^bDefined as number of positive and specific clones (no BSA binding) divided by the number of assayed clones (44).

^cDefined as unique sequences as determined by Sanger sequencing.

^dEstimated concentration of IgG in the HEK293 culture.

^ePercentage of protein in the peak of the proper size (~150 KDa) relative to the total mass as estimated by SEC-UPLC.

^fK_D estimated by BIAcore or ELISA; See Supplementary Material, Figure sF3.

^gTm of the second thermal transition corresponding to Fab unfolding as determined by Thermal-Shift Analysis.