Table 3. Batch analysis and stability data for PfAMA1-DiCo Drug Product.
| Test Parameter | Specification | At release | At T = 0 | T = 7 d 30°C | T = 3 m -20°C | T = 6 m -20°C | T = 9 m -20°C | T = 12 m -20°C | T = 18 m -20°C | T = 24 m -20°C | |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Appearance of lyophilised producta | White cake | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | |
| Appearance of reconstituted solutiona | Colourless to brownish, clear liquid without visible particles after reconstitution | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | |
| Moisture content | <4% w/w | <3% | <3% | 2.3% | 2.0% | 2.0% | 1.8% | 1.8% | 1.8% | 2.0% | |
| Dissolution ratea | <2 minutes | 30 sec | 30 sec | 29 sec | 7 sec | 17 sec | 8 sec | 7sec | 9 sec | 9 sec | |
| Sub-visible particles | ≤6000 particles of 10 μm in size or larger per container | 25 | 25 | 45 | n.d. | 13 | n.d. | 32 | n.d. | 16 | |
| ≤600 particles of 25 μm in size or larger per container | 2 | 2 | 4 | n.d. | 0 | n.d. | 3 | n.d. | 1 | ||
| Identity western blot | A principle band with relative molecular mass between 65 and 70 kDa | Complies | Complies | Complies | Complies | Complies | Complies | Complies | Complies | Complies | |
| Identity SDS-PAGE | A principle band with relative molecular mass between 65 and 70 kDa | Complies | Complies | Complies | Complies | Complies | Complies | Complies | Complies | Complies | |
| Identity and purity by SE-HPLC (%) | main peak >95% | 96.7 | 97.3 | 97.5 | 96.8 | 97.1 | 95.8 | 96.0 | 95.6 | 95.4 | |
| pH (-) | 6.3–7.3 | 6.5 | 6.5 | 6.5 | 6.4 | 6.4 | 6.8 | 6.4 | 6.8 | 6.4 | |
| Protein content by μBCA (μg/mL) | 79.5–129.5 μg/mL | 100.7 | 103.4 | 96.1 | 97.6 | 91.4 | 98.4 | 96.8 | 106.4 | 108.1 | |
| Total PfAMA1 DiCo content by SPR (μg/mL) | Report results | n.d. | 50.4 | 41.9 | 44.9 | 46.7 | 37.0 | 39.5 | 38.8 | 41.3 | |
| DiCo content by competition ELISA (μg/mL) | All three DiCos detectable in the reconstituted mixb Report resultsc | DiCo 1: | Pass | 35.1 | 32.1 | 50.3 | 26.4 | 32.1 | 45.1 | 40.8 | 32.1 |
| DiCo 2: | 34.4 | 36.5 | 32.9 | 35.8 | 37.0 | 39.6 | 30.4 | 34.4 | |||
| DiCo 3: | 25.1 | 35.9 | 31.5 | 21.2 | 30.7 | 28.3 | 26.5 | 28.5 | |||
| Potency (Alhydrogel) | Not less than 80% seroconversion | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | Pass | |
| Potency (GLA-SE) | Not less than 80% seroconversion | Pass | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | |
| Endotoxin | <30 IU/dose (50μg) (eq. to <36 IU/vial) | Pass | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | |
| Sterility | Sterile | Sterile | Sterile | n.d. | n.d | n.d. | nd | n.d. | n.d. | n.d. | |
| Abnormal toxicity | No weight loss, no abnormal reaction | Pass | n.d. | n.d. | n.d | n.d. | nd | n.d. | n.d. | n.d. | |
a. Reconstitution with 0.6 ml saline
b. Specification at release and for the stress test: 7d at 30°C
c. Specification for the real time stability study
n.d.: not done