Table IV.
Impact of Targeted Mutagenesis on Li33 Solubility
| Stability (°C) |
|||||
|---|---|---|---|---|---|
| Li33 Isotype | Solubilitya (mg/mL) | LINGO-1 binding, EC50 (nM) | TM1 | TM2 | SEC, % monomer |
| CDR targeted mutagenesis | |||||
| IgG1agly | 0.3 | 0.12 | 60 | 77 | >99 |
| IgG1aglyW94V/I57V | >10 | 0.15 | 59 | 74/82 | 99 |
| IgG1aglyW94V/I57S | >7 | 0.38 | 58 | 75/82 | 99 |
| IgG1aglyW94V/I57P | >8 | 0.07 | 58 | 75/83 | 99 |
| IgG1aglyW94V/I57T | >7 | 0.13 | 59 | 75/83 | 99 |
| IgG2wt | >50 | 0.23 | 69 | 76 | 96 |
| IgG2agly | 0.3 | 0.26 | 59 | 76 | 98 |
| IgG2-V234A/G237A | 5.6 | 0.19 | 69 | 76 | 95 |
| +W94V/W104Q/I57S | >50 | 2.9 | 67 | 74 | 95 |
| +W94V/W104Q/I57A | >50 | 2.4 | 69 | 73 | 95 |
| Glycosylation site insertion mutants | |||||
| IgG1wt | 0.9 | 0.12 | 69 | 76 | 99 |
| IgG1wt A120N | >50 | 0.10 | 69 | 75 | 95 |
| IgG1wt Q177N | 4.9 | 0.15 | 69 | 75 | 96 |
| IgG1wt T197N | >50 | 0.12 | 69 | 75 | 95 |
| IgG1wt K212S | 3.7 | 0.12 | 69 | 75 | 85 |
Samples were characterized for the limit in their solubility at 4°C, apparent binding affinity for LINGO-1, thermal stability, and aggregation state by SEC. All samples were analyzed at pH 7.0 in PBS.
a
Protocol for assessing solubility is described in the Materials And Methods section.