TABLE 3

Nanobody affinity measured using Alexa647-conjugated anti-His antibody on wild-type and mutated CXCR2 receptor CHO cell lines

Nanobody	pK_d of Nanobody Binding to CHO Cell Lines
Nanobody	CXCR2 WT	CXCR2-W15A	CXCR2-CCR9-ECL2	CXCR2-W15A:CXCR2-CCR9-ECL2
127D1 (class 1)	9.02 ± 0.06^a^/^a	No binding	9.18 ± 0.09^a^/^a	9.07 ± 0.10^a^/^a
163E3 (class 2)	8.44 ± 0.21^b^/^a	8.14 ± 0.18^a^/^a	No binding	8.11 ± 0.07^b^/^a
127D1-35GS-163E3 (biparatopic)	9.65 ± 0.14^c^/^a	8.72 ± 0.08^b^/^b	8.99 ± 0.05^a^/^b	9.71 ± 0.16^c^/^a

WT, wild type.
Values annotated with the same superscript letter are not statistically significantly different for a given receptor subtype across different nanobodies (values before /) or for a given nanobody across different receptors (values after /) (P < 0.05; Tukey’s honest significant difference test after two-way analysis of variance).