Table 2.
Comparison of final candidate formulation vs current dmLT formulation in terms of relative percent AB5 as measured by HIC, solution osmolality as well as physicochemical stability (agitation, thermal, freeze-thaw, chemical and conformational stability) properties as measured by MFI, OD350 assay, UV visible spectroscopy, Intact MS and DSC, respectively.
| Formulation | HIC | Osmolality | Agitation stability | Thermal stability (against aggregation) | F/T stability | Chemical stability glycation | Chemical stability oxidation | Conformational stability |
|---|---|---|---|---|---|---|---|---|
| Relative% (AB5) | (mOSm) | MFI (ΔTotal particles4h-0h > 2 µm, number/mL) | Temperature@OD350 = 0.1 unit (°C) | % Protein loss after 5FT | (Stressed at 40 °C for 7 days) | (Addition of 0, 1, 2.5 and 5 mM H2O2) | Tm values (DSC) | |
| 50 mM sodium phosphate, 50 mM NaCl, 10% w/v sucrose, 5 mM methionine, 0.1%v/v PS-80 pH 7.4 (Candidate formulation) | 66 ± 1 | 500 ± 7 | 8.9 ± 2.9 x 102 | 72.5 ± 0.2 | 0.0 ± 0.1 | No change in relative abundance of the glycated B-chain nor formation of additional glycated peaks | No oxidation detected at 1 and 2.5 mM H2O2 | Tm1 = 50.9 ± 0.2 Tm2 = 52.8 ± 0.1 Tm3 = 79.3 ± 0.1 Tm4 = 82.0 ± 0.1 |
| 42.7 mM sodium phosphate, 10.7 mM potassium phosphate, 82 mM NaCl, 5% lactose pH 7.4 (Current formulation) | 66 ± 1 | 408 ± 3 | 275.2 ± 11.8 x 102 | 74.4 ± 0.2 | 15.2 ± 0.3 | Increased glycation and formation of additional glycated peaks | Oxidation detected at 1, 2.5 and 5 mM H2O2 | Tm1 = 51.3 ± 0.2 Tm2 = 52.8 ± 0.1 Tm3 = 79.5 ± 0.1 Tm4 = 81.9 ± 0.1 |