- AAV production
Research Grade and NHP Grade AAV production
Nikebio delivers research-leading AAV packaging services to support your gene therapy projects. Our proprietary production platform, featuring optimized cell lines and enhanced RC and helper plasmids, maximizes virus yield. With years of expertise, we have perfected protocols to produce over 100 AAV serotypes with high titer, purity, potency, and consistency.
Focusing on the triple transient transfection system, we have developed a scalable upstream production process with suspension HEK293 cells and ultracentrifugation with iodixanol to meet the rapid preparation requirements of Research Grade AAV. As a result, our growing base of satisfied customers repeatedly relies on us for their AAV vector cloning and packaging needs.
Beyond Research grade AAV, we had developed a scalable downstream production process with Affinity and Anion exchange process, which meets the high quality of NHP grade AAV sample and easily scale up to pre-clinical studies. As shown in below, we successfully developed the manufacture process with affinity and anion exchange chromatography to purify AAV8 and AAV9 vector to guarantee the empty capsid rate <20% for NHP grade AAV.
| Research Grade | NHP Grade | |
|---|---|---|
| Application | Cell culture, small animal | Small animal or non-human primates |
| Scale | 2E+12~8E+14 GC | 1E+13~1E+16 |
| Lead time | Start from 12 business days | Start from 21 business days |
| QC test | dPCR, SDS-PAGE, Coomassie Blue Staining, endotoxin LAL | dPCR, SDS-PAGE, Coomassie Blue Staining, endotoxin LAL |
| Titration | dPCR, Titer meet requirements | dPCR, Titer meet requirements |
| AAV purity analysis | SDS-PAGE, Coomassie Blue Staining, Capsid band size meet reference | SDS-PAGE, Coomassie Blue Staining, Capsid band size meet reference, Capsid band size meet reference |
| Endotoxin Test | LAL, <10EU/mL | LAL, <1EU/ml (endotoxin removal may reach <0.5EU/ml) |
| Mycoplasma | - | qPCR, Negative |
| AAV Genome integrity | - | Alkaline gel electrophoresis |
| Empty Capsid | AUC, guaranteed empty capsid rate <30% for common serotypes | AUC, guaranteed empty capsid rate <20% for common serotypes |