Epitope mapping of CD66/CEACAM Mabs
| Mab . | δ N (CEACAM6 1-59 aa)* . | N (CEACAM6 1-122 aa)* . | CEA CEACAM1,3,6,8* . | CEACAM1 N-domain binding . | CEACAM1-1-Fc . | CEACAM1-3-Fc . | CEACAM1-4-Fc . | NCAM(D1-5)-Fc/ Muc 18-Fc† . |
|---|---|---|---|---|---|---|---|---|
| 26H7 | − | − | CEACAM1 | + | 1.498 ± 0.127 | 1.672 ± 0.068 | 1.659 ± 0.167 | 0.083 ± 0.003 |
| 5F4 | − | − | CEACAM1 | + | 1.774 ± 0.024 | 1.933 ± 0.090 | 1.931 ± 0.044 | 0.082 ± 0.001 |
| TEC-11 | − | − | CEACAM1 | − | 0.205 ± 0.005 | 0.188 ± 0.004 | 1.298 ± 0.121 | 0.191 ± 0.004† |
| 12-140-4‡ | nd | nd | CEACAM1 CEA | + | 1.560 ± 0.087 | 1.628 ± 0.253 | 1.731 ± 0.085 | 0.257 ± 0.018 |
| 4/3/17‡,2-153 | − | + | CEACAM1 CEA | + | 2.118 ± 0.106 | 2.178 ± 0.155 | 2.444 ± 0.101 | 0.416 ± 0.066† |
| b 7.8.5 | − | − | CEA | − | nd | 0.268 ± 0.019 | 0.267 ± 0.034 | 0.252 ± 0.007 |
| YG-C35D6 | − | − | CEACAM6 CEA | − | 0.360 ± 0.033 | 0.339 ± 0.040 | 0.356 ± 0.030 | 0.284 ± 0.010 |
| COL-4‡ | nd | nd | CEACAM1,3 CEA | + | 2.396 ± 0.60 | 2.319 ± 0.023 | 2.421 ± 0.122 | 0.310 ± 0.006 |
| F36-54 | − | − | CEACAM1,6 CEA | − | 0.187 ± 0.011 | 0.834 ± 0.031 | 0.999 ± 0.279 | 0.178 ± 0.016 |
| 34B1 | nd | nd | CEACAM1,3,6 CEA | + | 1.364 ± 0.117 | 1.507 ± 0.050 | 1.460 ± 0.089 | 0.800 ± 0.005 |
| YG-C28F2 | nd | nd | CEACAM1,3,6 CEA | + | 1.952 ± 0.275 | 1.872 ± 0.140 | 2.035 ± 0.164 | 0.271 ± 0.004 |
| D14HD11‡ | + | + | CEACAM1,3,6 CEA | + | 2.248 ± 0.168 | 2.175 ± 0.144 | 2.294 ± 0.163 | 0.221 ± 0.008 |
| b18.7.7‡ | + | + | CEACAM1,3,6 CEA | + | 1.795 ± 0.277 | 2.073 ± 0.346 | 1.822 ± 0.147 | 0.201 ± 0.058 |
| D11-AD11‡ | + | + | CEACAM1,3,6 CEA | + | 2.248 ± 0.168 | 2.175 ± 0.144 | 2.294 ± 0.163 | 0.221 ± 0.008 |
| HEA 81‡ | + | + | CEACAM1,3,6 CEA | + | 0.505 ± 0.120 | 0.577 ± 0.083 | 0.379 ± 0.062 | 0.206 ± 0.005† |
| B1.12-153 | − | + | CEACAM1,3,6 CEA | + | 0.870 ± 0.046 | 0.890 ± 0.126 | 1.051 ± 0.125 | 0.285 ± 0.002† |
| CLB-gran-102-155 | − | + | CEACAM1,3,6 CEA | + | 1.946 ± 0.389 | 2.071 ± 0.055 | 1.944 ± 0.086 | 0.206 ± 0.017 |
| F34-187‡,2-153 | − | + | CEACAM1,3,6 CEA | + | 2.019 ± 0.284 | 2.189 ± 0.148 | 2.135 ± 0.263 | 0.205 ± 0.006 |
| T84.1‡,2-155 | − | + | CEACAM1,3,6 CEA | + | 1.572 ± 0.138 | 1.701 ± 0.238 | 1.488 ± 0.317 | 0.235 ± 0.025 |
| B6.2 | nd | nd | CEACAM1,3,6 CEA | + | 1.913 ± 0.469 | 1.972 ± 0.335 | 2.325 ± 0.246 | 0.365 ± 0.039 |
| B1.13 | nd | nd | CEACAM1,3,6 CEA | + | 1.660 ± 0.050 | 1.698 ± 0.213 | 1.808 ± 0.194 | 0.253 ± 0.018 |
| YG-C94G7 | − | − | CEACAM1,6,8 CEA | − | 0.342 ± 0.037 | 1.469 ± 0.227 | 1.458 ± 0.062 | 0.279 ± 0.023 |
| 12-140-5 | − | − | CEACAM1,6,8 CEA | − | 0.444 ± 0.055 | 2.432 ± 0.033 | 2.357 ± 0.062 | 0.475 ± 0.051 |
| TET-2 | − | − | CEACAM1,6,8 CEA | − | 0.327 ± 0.023 | 2.041 ± 0.061 | 2.198 ± 0.262 | 0.289 ± 0.023 |
| mIgG1 | − | − | − | − | 0.147 ± 0.003 | 0.147 ± 0.003 | 0.150 ± 0.008 | 0.145 ± 0.005† |
| mIgG2a | − | − | − | − | 0.142 ± 0.002 | 0.135 ± 0.002 | 0.200 ± 0.010 | 0.206 ± 0.017† |
| mIgG2b | − | − | − | − | 0.199 ± 0.010 | 0.220 ± 0.019 | 0.144 ± 0.006 | 0.145 ± 0.011† |
| mIgG1 | − | − | − | − | 0.196 ± 0.016 | 0.223 ± 0.037 | 0.206 ± 0.008 | 0.184 ± 0.020 |
| mIgG2a | − | − | − | − | 0.188 ± 0.029 | 0.196 ± 0.004 | 0.201 ± 0.057 | 0.170 ± 0.009 |
| mIgG2b | − | − | − | − | 0.265 ± 0.111 | 0.156 ± 0.014 | 0.168 ± 0.012 | 0.166 ± 0.021 |
| mIgM | − | − | − | − | 0.171 ± 0.026 | 0.193 ± 0.026 | 0.201 ± 0.049 | 0.178 ± 0.028 |
| Mab . | δ N (CEACAM6 1-59 aa)* . | N (CEACAM6 1-122 aa)* . | CEA CEACAM1,3,6,8* . | CEACAM1 N-domain binding . | CEACAM1-1-Fc . | CEACAM1-3-Fc . | CEACAM1-4-Fc . | NCAM(D1-5)-Fc/ Muc 18-Fc† . |
|---|---|---|---|---|---|---|---|---|
| 26H7 | − | − | CEACAM1 | + | 1.498 ± 0.127 | 1.672 ± 0.068 | 1.659 ± 0.167 | 0.083 ± 0.003 |
| 5F4 | − | − | CEACAM1 | + | 1.774 ± 0.024 | 1.933 ± 0.090 | 1.931 ± 0.044 | 0.082 ± 0.001 |
| TEC-11 | − | − | CEACAM1 | − | 0.205 ± 0.005 | 0.188 ± 0.004 | 1.298 ± 0.121 | 0.191 ± 0.004† |
| 12-140-4‡ | nd | nd | CEACAM1 CEA | + | 1.560 ± 0.087 | 1.628 ± 0.253 | 1.731 ± 0.085 | 0.257 ± 0.018 |
| 4/3/17‡,2-153 | − | + | CEACAM1 CEA | + | 2.118 ± 0.106 | 2.178 ± 0.155 | 2.444 ± 0.101 | 0.416 ± 0.066† |
| b 7.8.5 | − | − | CEA | − | nd | 0.268 ± 0.019 | 0.267 ± 0.034 | 0.252 ± 0.007 |
| YG-C35D6 | − | − | CEACAM6 CEA | − | 0.360 ± 0.033 | 0.339 ± 0.040 | 0.356 ± 0.030 | 0.284 ± 0.010 |
| COL-4‡ | nd | nd | CEACAM1,3 CEA | + | 2.396 ± 0.60 | 2.319 ± 0.023 | 2.421 ± 0.122 | 0.310 ± 0.006 |
| F36-54 | − | − | CEACAM1,6 CEA | − | 0.187 ± 0.011 | 0.834 ± 0.031 | 0.999 ± 0.279 | 0.178 ± 0.016 |
| 34B1 | nd | nd | CEACAM1,3,6 CEA | + | 1.364 ± 0.117 | 1.507 ± 0.050 | 1.460 ± 0.089 | 0.800 ± 0.005 |
| YG-C28F2 | nd | nd | CEACAM1,3,6 CEA | + | 1.952 ± 0.275 | 1.872 ± 0.140 | 2.035 ± 0.164 | 0.271 ± 0.004 |
| D14HD11‡ | + | + | CEACAM1,3,6 CEA | + | 2.248 ± 0.168 | 2.175 ± 0.144 | 2.294 ± 0.163 | 0.221 ± 0.008 |
| b18.7.7‡ | + | + | CEACAM1,3,6 CEA | + | 1.795 ± 0.277 | 2.073 ± 0.346 | 1.822 ± 0.147 | 0.201 ± 0.058 |
| D11-AD11‡ | + | + | CEACAM1,3,6 CEA | + | 2.248 ± 0.168 | 2.175 ± 0.144 | 2.294 ± 0.163 | 0.221 ± 0.008 |
| HEA 81‡ | + | + | CEACAM1,3,6 CEA | + | 0.505 ± 0.120 | 0.577 ± 0.083 | 0.379 ± 0.062 | 0.206 ± 0.005† |
| B1.12-153 | − | + | CEACAM1,3,6 CEA | + | 0.870 ± 0.046 | 0.890 ± 0.126 | 1.051 ± 0.125 | 0.285 ± 0.002† |
| CLB-gran-102-155 | − | + | CEACAM1,3,6 CEA | + | 1.946 ± 0.389 | 2.071 ± 0.055 | 1.944 ± 0.086 | 0.206 ± 0.017 |
| F34-187‡,2-153 | − | + | CEACAM1,3,6 CEA | + | 2.019 ± 0.284 | 2.189 ± 0.148 | 2.135 ± 0.263 | 0.205 ± 0.006 |
| T84.1‡,2-155 | − | + | CEACAM1,3,6 CEA | + | 1.572 ± 0.138 | 1.701 ± 0.238 | 1.488 ± 0.317 | 0.235 ± 0.025 |
| B6.2 | nd | nd | CEACAM1,3,6 CEA | + | 1.913 ± 0.469 | 1.972 ± 0.335 | 2.325 ± 0.246 | 0.365 ± 0.039 |
| B1.13 | nd | nd | CEACAM1,3,6 CEA | + | 1.660 ± 0.050 | 1.698 ± 0.213 | 1.808 ± 0.194 | 0.253 ± 0.018 |
| YG-C94G7 | − | − | CEACAM1,6,8 CEA | − | 0.342 ± 0.037 | 1.469 ± 0.227 | 1.458 ± 0.062 | 0.279 ± 0.023 |
| 12-140-5 | − | − | CEACAM1,6,8 CEA | − | 0.444 ± 0.055 | 2.432 ± 0.033 | 2.357 ± 0.062 | 0.475 ± 0.051 |
| TET-2 | − | − | CEACAM1,6,8 CEA | − | 0.327 ± 0.023 | 2.041 ± 0.061 | 2.198 ± 0.262 | 0.289 ± 0.023 |
| mIgG1 | − | − | − | − | 0.147 ± 0.003 | 0.147 ± 0.003 | 0.150 ± 0.008 | 0.145 ± 0.005† |
| mIgG2a | − | − | − | − | 0.142 ± 0.002 | 0.135 ± 0.002 | 0.200 ± 0.010 | 0.206 ± 0.017† |
| mIgG2b | − | − | − | − | 0.199 ± 0.010 | 0.220 ± 0.019 | 0.144 ± 0.006 | 0.145 ± 0.011† |
| mIgG1 | − | − | − | − | 0.196 ± 0.016 | 0.223 ± 0.037 | 0.206 ± 0.008 | 0.184 ± 0.020 |
| mIgG2a | − | − | − | − | 0.188 ± 0.029 | 0.196 ± 0.004 | 0.201 ± 0.057 | 0.170 ± 0.009 |
| mIgG2b | − | − | − | − | 0.265 ± 0.111 | 0.156 ± 0.014 | 0.168 ± 0.012 | 0.166 ± 0.021 |
| mIgM | − | − | − | − | 0.171 ± 0.026 | 0.193 ± 0.026 | 0.201 ± 0.049 | 0.178 ± 0.028 |
+ indicates positive; −, nonreactive or negative; nd, not determined.
Summary from the Boston and Osaka Leucocyte Culture Conferences or after testing by flow cytometry on HeLa-CEA, CEACAM3L, CEACAM6, CEACAM8, and CHO-CEACAM1-4L, -4S and -1S.
Muc 18-Fc used in place of NCAM-Fc as a negative control; N domain covers 1-180 aa of CEACAM1-4L; δ N domain of CEACAM6 covers 1-59 aa; N domain of CEACAM6 covers 1-122 aa.
Mabs to the N-terminal domain that cross-block the binding of 125I-labeled 12-140-4 to CEACAM1 or CEA transfectants indicating similar or closely linked epitopes; other N-domain Mabs were not analyzed in cross-blocking experiments.
Does not cross-block CLB-gran/10.
Cross-blocks CLB-gran/10.