Supplementary Materials and Methods, Figures S1-5 and Tables S1-8 from Redesigning a Monospecific Anti-FGFR3 Antibody to Add Selectivity for FGFR2 and Expand Antitumor Activity

Supplementary Materials and Methods, Figures S1-5 and Tables S1-8. Supplementary Materials and Methods: More detailed description of Materials and Methods for experiments. Figure S1: Structural analysis of the FGFR3-IIIb and R3Mab complex. Figure S2: Sequence analysis of the CDR H2 of selected FGFR2-positive clones. Figure S3: Superimposition of crystal structures of FGFR2-D2:2B.1.3 and FGFR3-D2D3:R3Mab. Figure S4: in vitro and in vivo studies of the 2B.1 antibody variants using the breast cancer cell line MFM-223x2.2. Figure S5: FGFR2 and FGFR3 expressions in gastric cancer cell line SNU-16. Table S1: Library design for recruiting FGFR2 binding specificity for R3Mab. Table S2: Library design for removing FGFR4 binding specificity from 2B.1.3. Table S3: Sequence identities between human FGFR. Table S4: Bidning affinities of selected 2B.1 variants for FGFR2 and FGFR3. Table S5: X-ray crystallography data collection and refinement statistics. Table S6: Binding affinities of R3Mab and 2B.1.3 for the D2 alone and D2D3 constructs of FGFR2 and FGFR3. Table S7: Residue differences between FGFR2 and FGFR4 at the positions making potential contacts with 2B.1.3. Table S8: 2B.1.3 variants that retain FGFR2 binding and show diminished binding to FGFR4.