PackGene provides GMP-grade plasmid manufacturing services for cloning, amplification, and purification of plasmids to support subsequent GMP-grade AAV production as well as other applications. Our new segregated ISO-classified plasmid manufacturing suite (1000m²) is fully operational as of September 2021. We provide GMP-grade plasmid vector cloning, amplification, and purification services and cover large-scale fermentation, material isolation, full traceability documentation control, and comprehensive quality control assays.
PackGene’s plasmid production service is capable accepting existing plasmids provided by you for full scale plasmid production. Alternatively, we are able to reconstitute any existing plasmids by cloning your gene of interest into one of PackGene’s proprietary vector backbones to bolster production capacity. All cGMP plasmid production at PackGene employs complete upstream and downstream process development support.
PackGene’s cGMP fermentation ranges in scale from 50L to 200L with single use fermentation units. GMP plasmids are wholly separated from other plasmids and plasmid-bearing materials by physical barriers to maintain separate and clean production conditions.
After each production batch is completed we clean and maintenance the entire facility room. This includes thoroughly cleaning surfaces and all non-disposable equipment prior to the start of the next GMP plasmid production run. In addition to this cleaning regimen, and we use disposable materials for most critical procedures to significantly reduce the chance of contamination.
Bacteria bank generation
Quality control inspection and report
Single-use fermentation (50L to 200L)
Batch record, manufacturing summary report, CoA
Ultra-filtration & AKTA ready purification system
Plasmid Quality Control (or customized based on specific process)
| Category | Items | Method |
| General Characterization | Appearance | Visual inspection |
| pH | pH meter | |
| Identity | Restriction Digest | Agarose gel electrophoresis |
| Plasmid Sequence | Sanger Sequencing | |
| Content | DNA Concentration | UV spectrophotometry(A260) |
| Purity | UV Purity | A260/A280 ratio |
| DNA Homogeneity | HPLC | |
| Impurity | Residual Host Cell DNA | qPCR |
| Residual Host Cell RNA | Agarose gel electrophoresis | |
| Residual Host Cell Protein | ELISA | |
| Residual Kan | ELISA | |
| Safety | Endotoxin | LAL |
| Sterility Test | Direct Inoculation Method |
Optimization
Process developed with Quality by Design (QbD) concepts and optimization
Speed
GMP-compliant pilot production cycle can as fast as 1-2 months
Cleanliness
Meticulously cleaned environment in accordance with C+A standards
Rigor
Isolation
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.
CDMO Plasmid Flyer
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