Table 2.
Developability of CPP-Abs depending on selected insertion positions.
| CPP | Position | Ab concentration in culture supernatant (mg/L) | SEC after affinity purification | Purity after 2 step purification (%) | Purification yield (%) | LC-MS |
|---|---|---|---|---|---|---|
| no | Not applicable | 141 | 1 peak | 97 | 80 | Expected mass |
| Pep-1 | N-ter LC (LCN) | 2 | nd | nd | nd | nd |
| Pep-1 | C-ter LC (LCC) | 145 | 1 peak | 96 | 80 | Expected mass |
| Pep-1 | N-ter HC (HCN) | 1 | nd | nd | nd | nd |
| Pep-1 | C-ter HC (HCC) | 113 | 6 peaks | 94 | 27 | Expected mass + others |
| Pep-1 | before hinge (BH) | 132 | 1 peak | 97 | 63 | Expected mass |
| Pep-1 | after hinge (AH) | 120 | 1 peak | 95 | 43 | Expected mass |
| TAT | N-ter LC (LCN) | 4 | nd | nd | nd | nd |
| TAT | C-ter LC (LCC) | 174 | 2 peaks | 74 | 43 | truncated LC in TAT sequence |
| TAT | N-ter HC (HCN) | 5 | nd | nd | nd | nd |
| TAT | C-ter HC (HCC) | 190 | 2 peaks | 88 | 20 | truncated LC in TAT sequence |
| TAT | before hinge (BH) | 156 | 6 peaks | 75 | 26 | truncated HC in TAT sequence |
| TAT | after hinge (AH) | 156 | 3 peaks | 67 | 41 | truncated HC in TAT sequence |
Nd: not determined. SEC: size exclusion chromatography.