Table 1.

BIAcore SPR and DLS results summary table for all variants. Tabulated are SPR KD (nM) values, the ratio of the measured KD and the HC-F104 reference KD (KD/KD_ref), and kD (mL/g) values obtained by DLS. The reference F104 is highlighted gray. All LC-Y30 and LC-F92 variants are site-directed mutants of the F104 variant. The single F104W variant with comparable BIAcore SPR KD to the reference is highlighted green. Variants with 10–20× differences in binding relative to the variant are highlighted yellow, and those with greater than 20-fold difference in SPR binding or non-binding (NB) are highlighted red. kD (mL/g) measured by DLS is color coded in terms of degree of self-association (greater than −20 mL/g, green; −20 to −29 mL/g, yellow; −30 mL/g or less, red). “NA” denotes no measurement.

Variant	KD (nM)	KD/KDref	kD (mL/g)
F104	0.35	1	−44.6
F104W	0.16	0.44	−15.3
F104I	108	305	−36.9
F104H	1372	3879	−38.5
F104D	NB	NB	−10.9
F104K	NB	NB	−4.8
F104E	NB	NB	−18.1
F104S	NB	NB	−20.1
F104G	NB	NB	−41.6
F104R	NB	NB	−10.7
Y30R	4.36	12.3	−18.9
Y30H	5.5	15.6	NA
Y30N	45.5	129	NA
Y30D	60.9	172	−7.9
Y30Q	82.6	234	NA
Y30G	170	481	−42.9
F92W	5.51	15.6	−37.4
F92H	35.1	99.2	−20.7
F92V	38.7	109	−21.4
F92R	50.2	142	−10.8
F92S	62.7	177	−31
F92G	142	402	−31.3