AAV cGMP Manufacturing
As gene therapy continues to advance and develop rapidly, the need for large-scale production of GMP-grade AAV gene therapy specialty services that meet regulatory filing standards has become urgent, from IIT (PI initiated clinical trials), IND filings and clinical Phase I-III clinical trials to large-scale commercial production of large quantities of GMP-grade AAV. manufacturing, processing, batch testing and IND pharmacology study writing services.
Utilizing our intellectual properties, PackGene provides mass-production services of GMP-compliant AAV, as well as GMP production line setting-up, processing and assay transfer, IND pharmacological documentation, ranging from the upstream rAAV generation to the downstream isolation and purification technologies. We also ensure AAV manufacturing services comply with GMPs and all applicable regulatory and required standards for IIT and IND applications.
Note: PackGene’s FDA and EMA compliant AAV GMP manufacturing facility （~2000m²） start to begin production in 2021. For more information, please contact us: bd@packgene.com
Interested in our AAV services? Please contact us for a free consultation or submit a request for a quotation.
AAV Manufacturing Process
PackGene has developed core processing technology that multiply the single-batch AAV output/yield and reduce the input cost, giving our customers a leading edge.
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 |
AAV Manufacturing Facilities
Our GMP manufacturing site fully adopts “disposable technology” and advanced design concepts, and is equipped with a clean environment according to the quality by design (QbD) concept, with physical isolation and full-exclusion systems to segregate the different functional workshops, of which the GMP workshop is only used for pilot testing of AAV carriers The GMP workshop is only used for pilot testing or large-scale production of AAV carriers, which completely avoids the limitations of multi-species sharing or cross-contamination commonly found in traditional CDMO platforms.
The upstream production line of the GMP site is equipped with the leading iCellis 500 disposable bioreactor system (up to 500m2 wall area), STR200T 200L disposable reactor, Xpansion disposable reactor, XDR 50 MO 50L disposable reactor, and several wave-shaped bioreactors to meet the needs of large-scale wall and suspension culture production of AAV. AAV production. The downstream line is equipped with ÄKTA chromatographic purification and ultrafiltration systems. Based on years of experience in large-scale production, single batch AAV yields of 1E+15-17GC or vg can be achieved, allowing for a shortened pilot production cycle of AAV under GMP standards to be completed within 1-2 months. Paijin’s core process technology route can help partners to significantly increase single batch yields and reduce input costs, gaining valuable speed advantages for customers and accelerating registration filings and clinical level production of gene therapy speciality services.
The purpose of the production final generation cell assay is to confirm their suitability throughout the production process and whether the cells are mutated or produce carcinogenic substances. Cells are resuscitated and passaged from the working cell bank according to the production process route, transferred to the bioreactor for passaged digestion, cultured according to the proposed process, and then digested for lyophilization or sent for testing at the end of bioreactor production. The generation of cell resuscitation and passaging methods are carried out according to the production process SOP.
PackGene facility includes iCellis500 (~500m2 adhesion area), STR50/200/500T with the single-use technology that can flexibly conduct both GMP AAV upstream production by adherent and suspension culture.
Our advantages
- Designed with QbD concept and optimization
- Clean environment in accordance with C+A standards
- Reaching the maximum AAV yield up to 1E+15GC-1E+17 GC/batch
- Isolated vector producing line to completely avoid cross-contamination between different vectors
- GMP-compliant pilot production cycle can be shortened to 1-2 months
- Single-use technology and water supply to ensure batch cleaning verification
