TABLE 2.
Summary of On-target and Near Off-target Activity of HPV E711–19 TCR and Examples of Best Primary CARs in Jurkat:T2 Assays
| Jurkat:T2 Assay | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Constructs | Construct Format or Source | Test Peptide Name | Test Peptide Sequence | On/Off Target | EC50 (μM) | ECminimum (μM) | Eminimum (RLU) | Emaximum (RLU) | Selectivity |
| C882 | TCR | HPV E711–19 | YMLDLQPET | On | 0.00005 | 0.000008 | 900 | 14,000 | NA |
| ZFN236819–828 | MLDLEPQHVV | Off | >100 | NA | 740 | NA | >2,000,000 | ||
| ZFN236819–827 | MLDLEPQHV | Off | >100 | NA | 900 | NA | >2,000,000 | ||
| SH3GLB1242–252 | QYMLDLQKQL | Off | >100 | NA | 900 | NA | >2,000,000 | ||
| SH3GLB1243–252 | YMLDLQKQL | Off | >100 | NA | 900 | NA | >2,000,000 | ||
| C1007 | Primary binder CAR | HPV E711–19 | YMLDLQPET | On | 1.7 | 0.137 | 16,000 | 52,000 | NA |
| ZFN236819–828 | MLDLEPQHVV | Off | 35 | ND | 17,000 | 39,000 | 21 | ||
| ZFN236819–827 | MLDLEPQHV | Off | >100 | NA | NA | NA | >59 | ||
| SH3GLB1242–252 | QYMLDLQKQL | Off | >100 | NA | NA | NA | >59 | ||
| SH3GLB1243–252 | YMLDLQKQL | Off | >100 | NA | NA | NA | >59 | ||
| C996 | Primary binder CAR | HPV E711–19 | YMLDLQPET | On | 0.07 | 0.005 | 6806 | 27,604 | NA |
| ZFN236819–828 | MLDLEPQHVV | Off | >100 | NA | NA | NA | >1428 | ||
| ZFN236819–827 | MLDLEPQHV | Off | >100 | NA | NA | NA | >1428 | ||
| ZFN236818–827 | AMLDLEPQHV | Off | 13.2 | ND | 2365 | 3705 | 189 | ||
| SH3GLB1242–252 | QYMLDLQKQL | Off | 13.7 | ND | 1994 | 6108 | 196 | ||
| SH3GLB1243–252 | YMLDLQKQL | Off | 6.5 | ND | 2245 | 7111 | 93 | ||
C996 is the parent of C1800 and C1806 shown in Supplementary Figure 4A and Supplementary Table 3 (Supplemental Digital Content 1, http://links.lww.com/JIT/A623).
CAR indicates chimeric antigen receptor; HPV, human papillomavirus; NA, not applicable; ND, not determined; RLU, relative luminescence unit; TCR, T-cell receptor.