Overview
- Cell Bank Generation
- Glass Bioreactor (1-10L) and 50L single use bioreactor for suspension culture；
- Pall Nano 4m² adherent bioreactor（0.53-4 m²）
- CoA
- Ultra-centrifugation (optional) & AKTA Pure purification system
- AAV process development
- IND filing document preparation
AAV Quality Control (or customized based on specific process)
| Specification | Assay Methods | |
| Identity | GOI Sequence | Sanger Sequence |
| AAV Capsid | SDS PAGE/MS | |
| Purity
|
HPLC Purity | HPLC |
| UV Purity | A260/A280 | |
| %Empty Capsids | AEX HPLC | |
| TEM | ||
| AUC | ||
| Aggregation | DLS | |
| Potency & Content | Capsid Titer | ELISA |
| Genome Titer | ddPCR | |
| Infectious Titer | TCID50 | |
| Protein Expression | In Vitro assay | |
| Impurity | Residual Host Cell DNA | qPCR |
| Residual E1A | qPCR | |
| HCP Residue | ELISA | |
| BSA Residue | ELISA | |
| Residual Plasmid | ddPCR | |
| Residual Nuclease | ELISA | |
| Residual Iodixanol/CsCl | HPLC | |
| Residual Ligand | ELISA | |
| Safety | Mycoplasma | qPCR/Mycoplasma culture /DNA staining |
| rcAAV | Cell Assay+qPCR | |
| Bacterial endotoxin | LAL | |
| Sterility | Direct Inoculation Method/Membrane filtration | |
| Adventitious agent | Cell Culture | |
| Abnormal toxicity | Animal test | |
| General Characterization | Appearance | Visual Inspection |
| Content | Minimum Fill | |
| pH | pH Method | |
| Osmolality | Osmometers | |
| Particulate matter | Light Obscuration Particle Count test |
Flexibility
Both suspension and adherent culture process are available at multiple scales of production.
Professional
PackGene has a professional AAV team with more than 10 years of experience.
Advanced equipment
Pall Nano bioreactor suitable for adherent culture while it’s the scale-down model icellis 500, which is available at PackGene’s cGMP production services. Amber 250 ultra-scale down model used for high through-put process development available.
Efficiency
PackGene’s experienced team, advanced equipment, and streamlined processes ensure that your project will be completed in a timely manner.
Resources
Are pH measurements required, and is a large amount of sample wasted to carry out pH measurements?
Measurement of pH is a mandatory for the release of rAAV Fast Service deliverables. A micro pH electrode may be used to save sample and thus the required sample volume to perform pH measurements is only ~15uL-100uL.
What is loading?
In accordance with the Pharmacopoeia General Rules 0942, we use the minimum filling quantity inspection method for detecting sample loading quantity.
How to interpret A260/A280 value?
A260/A280 is the ratio of sample absorbance measured at wavelengths of 260nm and 280nm. This measure is commonly thought to represent the ratio of DNA to protein in a sample. For rAAV, A260/A280 can used as a measure of the full to empty shell rate and to identify protein contamination. Low A260/A280 levels may suggest that the empty shell rate is high. Alternatively, high A260/A280 may suggest that the sample has been contaminated with proteins that are not incorporated into the AAV capsid shell. The greatest advantages of this measure are its convenience and speed.
What tests are performed to differentiate rAAV capsid proteins from specific protein impurities?
SDS-PAGE is used to identify rAAV capsid proteins. In addition, SDS-PAGE can be used to directly identify specific protein impurities including the presence of host proteins, BSA, or degraded AAV capsid proteins.
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