Table I.
Impact of Li33 IgG Frameworks on Solubility
| Stability (°C) |
|||||
|---|---|---|---|---|---|
| Li33 isotype | Solubilitya (mg/mL) | LINGO-1 binding, EC50 (nM) | TM1 | TM2 | SEC, % monomer |
| IgG1wt pH 7.0 | 0.9 | 0.12 | 69 | 76 | 99 |
| IgG1agly pH 7.0 | 0.3 | 0.12 | 60 | 77 | >99 |
| IgG4wt pH 7.0 | >30 | 0.35 | 64 | 72 | 98 |
| IgG4agly pH 7.0 | 0.3 | 0.37 | 56 | 73 | 95 |
| IgG2wt pH 7.0 | >50 | 0.23 | 69 | 76 | 96 |
| IgG2agly pH 7.0 | 0.2 | 0.26 | 59 | 76 | 98 |
| IgG2-V234A/G237A pH 7.0 | 5.6 | 0.19 | 69 | 76 | 95 |
| IgG1 Fab2 pH 7.0 | 0.3 | 0.10 | – | 77 | 98 |
| IgG2 Fab2 pH 7.0 | >50 | 0.39 | – | 77 | 98 |
| IgG1 Fab pH 7.0 | >50 | 0.68 | – | 76 | 95 |
| IgG1agly reduced pH 7.0 | >40 | 0.12 | 55 | 75 | 98 |
| IgG1wt reduced pH 7.0 | >50 | 0.15 | 63 | 75 | 98 |
| IgG1wt pH 6.5 | 1.7 | 0.10 | 69 | 77 | |
| IgG1wt pH 6.0 | 2.4 | 0.10 | 69 | 78 | |
| IgG1wt pH 5.5 | 30 | 0.16 | 66 | 81 | |
| IgG1wt pH 5.0 | >50 | 0.45 | 66 | 81 | |
| IgG1wt pH 4.5 | >50 | 2.1 | 62 | 82 | |
| IgG1wt pH 4.0 | >50 | 16 | 54 | 78 | |
| IgG1wt pH 3.5 | >50 | 34 | 46/66 | 74 | |
| IgG1 wt pH 3.0 | >50 | >100 | 34/52 | 72 | |
Li33 Mabs were characterized for the limit in their solubility at 4°C, apparent binding affinity for LINGO-1, thermal stability, and aggregation state by SEC (samples loaded at 300 μg/mL). Samples at pH 7.0 were prepared in 20 mM sodium phosphate, 150 mM NaCl. Samples at other pHs were prepared in 10 mM sodium citrate, 150 mM NaCl.
a
The protocol for assessing solubility is described in the Materials and Methods section.