Table 1.

Characteristics of clones isolated from directed evolution of V_H-V_L M8 and M8V_L FAPs.

Clone name	Format	Fluorescence enhancement (YSD)*	Soluble KD or KD2/α	ϕ f	Mutations in sequence different from wild type (VH-VL M8 or M8VL)
M8	VH-VL	1.0	ND
L9	VH-VL	5.7X	ND		FH29S, WH36C, AH93V, IH113iT
Q9	VL	3.8X	1.3±0.3×10−15 M2	0.19	QL1R, TL14I,D L30G,QL37R, SL55P, FL62L, SL80P
J8	VL-VL	6.4X	<0.1 nM†		VL#1: DL85N, KL103T, LL107S, IL108iT VL#2: SL9P, SL32P
M8VL	VL	3.7X	2.5±0.7×10 15 M2	0.71
M8VL SL55P	VL	8.2X	1.0±0.5×10−16 M2	0.58	SL55P

^*Fluorescence enhancement is calculated as DIR fluorescence signal normalized for cell surface expression for each individual clone, divided by the equivalent value for the wild type parent (V_H-V_L M8). The concentration of Q9 was 33 nM, J8 was 9 nM, M8V_L was 10 nM, M8V_LS^L55P was 135 nM.

^†Data were equally well fit to a wide range of K_d values less than 0.11 nM.