Table 3.
Epitope mapping of recombinant IgGs (converted from scFv) and murine monoclonal antibodies
| ELISA1 |
epitope mapping CXCR2 N-terminus5 |
||
|---|---|---|---|
| [mAb](ng/ml)2 |
|||
| IgG | Nterm-N3 | Nterm-C4 | MEDFNMESDSFEDFWKGEDLSNYSYSSTLPPFLLDAAPCEPESLEINK |
| 4138.51 | 12 | > 1000 | ESDSFEDFWK |
| 4081.73 | 6 | > 1000 | FEDFWK SSTLPPFLLDA |
| 4138.73 | 4 | > 1000 | DSFEDFWK |
| 4138.06 | 8 | > 1000 | FNMESDSFEDFWK YSSTLPPFLLD |
| 4138.39 | 9 | > 1000 | FEDFWK |
| 4139.86 | 37 | > 1000 | MESDSFEDFWKGEDLSN |
| 4138.38 | 8 | > 1000 | FEDFWKGEDLSNYSYSSTLPPFLLDA |
| 4138.18 | 22 | > 1000 | ESDSFEDFWKGE LSNYSYSSTLPPFLLD |
|
murine mAb |
|
|
|
| mAb 27 | > 1000 | 4 | FEDFWKGEDLSN LPPFLLDAAP |
| mAb 5 | > 1000 | 3 | FEDFWKGEDL LPPFLLDAA |
| mAb 9 | > 1000 | 3 | FEDFWKGEDL PPFLLDAA |
| mAb 7 | > 1000 | 4 | LPPFLLDAAPCE |
| mAb 17 | 7 | 629 | DSFEDFWKGEDL PPFLLDAAP |
| mAb 19 | > 1000 | 276 | PPFLLDAA |
| mAb 21 | 6 | > 1000 | PPFLLDAA |
1The binding concentration of all listed monoclonal antibodies was > 1000 ng/ml for linear ECL1 (biotin-Ahx-GGAASKVNGWIFGTFLAK-NH2), cyclic ECL1 (biotin-Ahx -CASKVNGWIFGTFL C(Acm)KC-NH2), linear ECL3 (biotin-Ahx -GGDTLMRTQVIQETAERRNHIDR-NH2) and cyclic ECL3 (biotin-Ahx -GGhCDTLMRTQVIQETAERRNHIDRGhC-NH2) representing peptides. The cyclic peptides were cyclized by CLIPS™ moieties.
2[mAb] (ng/ml) at 4-fold background OD.
3Nterm-N = linear N-terminal part of the CXCR2 N-terminus: biotine-Ahx-GGMEDFNMESDSFEDFWKGEDLSNYSYSS-NH2 (Ahx = alpha hexanoic acid).
4Nterm-C = linear C-terminal part of the CXCR2 N-terminus: biotine-Ahx-GGSYSSTLPPFLLDAAPAEPESLEINK-NH2.
5Epitope mapping included 51 overlapping 16-mers peptide array comprising the N-terminus, ECL1 (data not shown), ECL2 (data not shown), and ECL3 (data not shown) domains of hCXCR2. Dominant epitopes are indicated in bold, weak epitopes in regular.