AAV Capsid Libraries Production
Producing high-quality and diverse AAV capsid libraries, can be technically challenging. AAV library variants are prone to cross-packaging and capsid mosaic formation when produced at high plasmid levels. Producing an AAV capsid library in low copy number can help reduce the cross packaging and capsid mosaic formation. However, reduce the library plasmid copy number during the transfection would introduce another challenge, generating a large number of AAV vectors efficiently and reproducibly. By using the mini-pHelper-Rep plasmid and high efficiency suspension HEK 293T cell lines, AAVnerGene was able to make AAV capsid libraries production more accessible and affordable for researchers and biotech companies.
Producing high-quality, diverse AAV capsid libraries presents technical challenges. One major hurdle is ensuring each vector in the library has a unique DNA sequence. Errors or duplications in the transgene region can result in biased or incomplete library representation.
Key Challenges in AAV Library Production:
- Cross-Packaging & Mosaicism:
When producing AAV libraries, cross-packaging and mosaicism can occur. This happens when capsid particles contain genomes and capsid monomers from different library members, leading to inconsistencies. Producing the library at a high plasmid level exacerbates this issue, while reducing plasmid copy number can minimize cross-packaging but creates inefficiencies in vector generation. - Balancing Yield & Quality:
Lowering plasmid copy numbers during transfection reduces cross-packaging and mosaicism, but it presents the challenge of generating sufficient AAV vectors efficiently and reproducibly. Innovative methods to improve AAV library yield while maintaining low cross-packaging are needed to create larger libraries with greater diversity. This diversity is crucial for identifying effective AAV capsids for gene therapy applications.
AAVnerGene’s Solutions:
- Mini-pHelper-Rep Plasmid: At AAVnerGene, we’ve developed a mini-pHelper plasmid, which replaces the traditional Ad helper plasmid. Its compact size improves transfection efficiency and lowers production costs. To streamline the process further, we created the mini-pHelper-Rep plasmid, which integrates the Rep gene into the mini-pHelper plasmid. By co-transfecting this plasmid with the AAV library plasmids, AAV libraries can be packaged more efficiently.
- High-Efficiency Suspension HEK 293T Cells: Using these specially optimized cells along with the mini-pHelper-Rep plasmid, AAVnerGene has made AAV library production more scalable, accessible, and cost-effective for both academic researchers and biotech companies. This advancement allows for the creation of large, diverse libraries while maintaining high-quality production standards.
Design of mini-pHelper-Rep Plasmid
AAV Capsid Libraries Generated from AAVnerGene
High Complexity:
High Diversity:
Low Cross-packaging:
AAV Capsid Library Production Services
By using the mini-pHelper-Rep plasmid and high efficiency suspension HEK 293T cell lines, AAVnerGene was able to make AAV capsid libraries production more accessible and affordable for researchers and biotech companies.
Scale | Packaging Conditions | Price | Turnaround |
2.00E+12 | 1000 copies/cell | $11,800 | 2-3 weeks |
2.00E+12 | 200 copies/cell | $19,800 | 2-3 weeks |
2.00E+12 | 100 copies/cell | $29,800 | 2-3 weeks |
2.00E+12 | <50 copies/cell | Please call to discuss | |
2.00E+13 | 1000 copies/cell | $48,800 | 3-4 weeks |
2.00E+13 | 200 copies/cell | $98,000 | 3-4 weeks |
2.00E+13 | 100 copies/cell | $148,800 | 3-4 weeks |
2.00E+13 | <50 copies/cell | Please call to discuss | |
2.00E+14 | Please call to discuss | Please call to discuss | |
AAV Capsid Library Related QC Services
Quality control (QC) is a critical step in ensuring that AAV libraries are suitable for downstream applications. In addition to regular AAV characterization, AAVnerGene provides Next-Generation Sequencing (NGS) services to verify the complexity, integrity, and overall quality of the AAV capsid library. This ensures that the library maintains its diversity and is ready for effective screening.