Table 1. Biophysical characteristics of V_Ls.

VL	Subgroup	Mfor (kDa)	MMALS (kDa)a	Expression yield (mg)b	KD (µM)	VL:protein Lc	Tm (°C)	ΔTm (°C)	TRE (%)d		GI protease resistance (%)e
									4 µM	20 µM	Trypsin	Chymotrypsin	Pepsin
HVLP324	Vκ1	13.83	13.84 ± 0.76	2.5, 7	0.2, 0.07f	7:1	66.1	7.3	90	78	34	100	22
HVLP324S		13.87	14.68 ± 0.58	0.5, 1.1	0.1, 0.06f	7:1	73.4		NDg	NDg	32	36	46
HVLP325	Vκ3	13.61	13.5 ± 0.63	0.5, 6.2	1	3:1	68.5	14	94	65	67	76	34
HVLP325S		13.66	13.35 ± 0.64	2.2, 3.1	1	3:1	82.5		NDg	NDg	81	93	100
HVLP335	Vκ3	13.90	15.26 ± 0.50	73.5	2	3:1	61.7	17.3	92	95	2	59	0
HVLP335S		13.94	16.8 ± 0.47	5.5	2	3:1	79.0		NDg	NDg	0	32	62
HVLP342	Vκ1	13.95	14.22 ± 0.64	1, 7.7	0.6, 0.04f	7:1	58.4	5.4	92	70	25	73	3
HVLP342S		13.99	17.02 ± 0.40	1.7, 10.8	0.2, 0.05f	7:1	63.8		NDg	NDg	5	40	5
HVLP351	Vκ3	13.85	14.60 ± 0.61	1.2, 8.9	2	2:1	62.0	9.9	87	65	71	100	31
HVLP351S		13.89	14.61 ± 0.53	1.9, 4.8	0.7	2:1	71.9		NDg	NDg	100	99	94
HVLP364	Vκ3	13.95	14.20 ± 0.58	0.3, 77	3	3:1	57.0	15.3	100	92	32	58	0
HVLP364Sh		13.97	14.98 ± 0.43	4.7	NDg	NDg	72.3		NDg	NDg	0	12	0
HVLP3103	Vκ3	13.85	14.29 ± 0.64	0.7, 19	1	3:1	65.7	10.7	94	92	81	90	0
HVLP3103S		13.89	14.05 ± 0.68	6.5	1	3:1	76.4		NDg	NDg	47	80	89
HVLP389	Vλ1	13.69	15.12 ± 0.50	3, 16.7	1	1:1	51.9	14.4	91	91	15	74	0
HVLP389S		13.72	13.65 ± 0.68	6.5	1	1:1	66.3		NDg	NDg	64	90	11

^a Mean ± SEM; ^bExpression yield values are per liter of bacterial culture. Two expression yield values correspond to two independent expression trials; ^cStoichiometry of V_L-protein L binding; ^dThermal refolding efficiency at 4 and 20 µM V_L concentrations, respectively; ^ePercentage proteolytic resistance values are at protease concentrations of 10 µg/mL (see also Fig. 6E); ^fSmaller K_D values correspond to the binding of HVLP324, HVLP324S, HVLP342, and HVLP342S to the high affinity sites on protein L; ^gND, not determined; ^hHVLP364S additionally gave a significant second peak (Fig. 3A) with a smaller elusion volume and a M_MALS of 26.21 ± 0.47 kDa.