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. 2011 Oct 31;287(2):1458–1467. doi: 10.1074/jbc.M111.253658

FIGURE 1.

FIGURE 1.

Determination of potential residues involved in chemokine interaction with dual-specific scFvs using binding specificity against CXCL10 and CXCL9 proteins from different species. A–C, specific binding of E7, J9, P8, F13, C1, and J5 dual-specific scFvs to a panel of immobilized CXCL10 (A), CXCL9 (B), and CXCL11 (C) chemokines from different species was assessed in an ELISA. The indicated NusA-fusion chemokines were captured and then incubated with scFvs. Coating was controlled using a specific anti-NusA mAb, and NusA protein was also added to the assay as a negative control (data not shown). Results are expressed as mean ± S.D. of duplicates. D and E, sequence alignments of mature CXCL10 (D) and CXCL9 (E) chemokine amino acid sequences from different species. Residues that are identical for human and cynomolgus but different from rabbit CXCL10 (D) or residues that are identical for human and mouse but different from cynomolgus CXCL9 (E) are shaded black. These residues were targeted for mutagenesis and substituted with the corresponding human residue. Numbering was done according to the template sequence, rabbit CXCL10 or cynomolgus CXCL9. hum10, human CXCL10; mou10, mouse CXCL10; cyn10, cynomolgus CXCL10; rat10, rat CXCL10; rab10, rabbit CXCL10; hum9, human CXCL9; mou9, mouse CXCL9; cyn9, cynomolgus CXCL9.