Gene Therapy AAV Packaging Services in Southeast Asia

Overcoming the Challenges of NHP Studies

Non-human primate (NHP) studies are essential for evaluating the safety and efficacy of gene therapies. However, these studies can be complex and costly from animal care, housing and maintenance. PackGene offers high-quality NHP-grade AAV vectors to streamline your research and improve your outcomes.

Traditional challenges include:

• High Cost: NHPs are expensive to acquire and maintain.
• Regulatory Hurdles: Strict regulations and ethical considerations can delay study timelines. (Requires significant amount of paperwork and approvals)
• Variable AAV Quality: Low-quality AAV vectors can lead to inconsistent results and compromised efficacy.
• Immune Response: NHPs can mount immune responses to AAV vectors, reducing their therapeutic potential.

Why Choose PackGene’s NHP-Grade AAV Vectors?

Our high-quality NHP-grade AAV vectors are designed to address these challenges and streamline your NHP studies:

• Tailored Production: We offer a comprehensive range of AAV serotypes, including popular choices like 1, 3b, 5, 7, 8, 9, and Rh10. For less common or experimental serotypes, we can conduct small-scale pilot studies to optimize production parameters and ensure consistent yields.
• Rigorous Quality Control: Our stringent quality control measures guarantee the purity, potency, and safety of our NHP-grade AAV vectors. We perform a comprehensive battery of tests, including:
• Enhanced Efficacy & Genome Integrity: Achieve higher levels of gene expression and therapeutic outcomes. Ensuring the correct sequence and structure of the AAV genome.
• Reduced Immune Response: Minimize off-target effects and improve safety by controlling empty capsid rate and endotoxins to minimise the presence of non-functional particles. The absence of microbial contaminants is ensured.
• Improved Reproducibility: Consistent results across different studies.
• Faster Time to Market: Accelerate your research and development timeline.

Our NHP-Grade AAV Production Process:

1. Vector Design and Cloning: We design and construct your gene of interest into AAV plasmids.
2. AAV Vector Production: We utilize state-of-the-art techniques to produce high-titer NHP-grade AAV vectors.
3. Purification and Concentration: We employ a multi-step purification process to remove cellular debris and other impurities.
4. Rigorous Quality Control: Our comprehensive QC measures ensure purity, potency, and safety.
5. Sterile Filtration and Aliquoting: We sterile filter the AAV vectors and aliquot them into appropriate volumes for storage and use.

Quality Control Standards of AAV Packaging – NHP Grade specifications

Plasmids used for AAV NHP Grade exceeds industrial standards

To ensure the fidelity of the gene of interest and efficient AAV packaging, the plasmids utilized for NHP grade AAV packaging undergo strict QC. The basic standards consist of requirements for appearance, A260:280 ratio, homogeneity (supercoil >90±10%), restriction analysis, residual RNA and E. coli DNA, endotoxin (≤0.01 EU/µg), and ITR and GOI Sanger sequencing, which are crucial for ensuring the efficient delivery of therapeutic genes via AAV.

In addition to the basic standards, there are also advanced requirements that are not included in the default package and are available upon request with additional cost. These advanced requirements include residual host protein, advanced endotoxin removal, bioburden, mycoplasma contamination, ITR Nanopore sequencing, animal-free production, material archiving, pH (7.5~8.0), residual Kan, USP sterility, dedicated purification resin or filtration membrane, etc.

Performance

The test methods described are necessary to ensure the quality and purity of AAV (adeno-associated virus) produced by PackGene, especially for its use in non-human primates (NHP). The different test methods evaluate various aspects of AAV quality, including its genome integrity, capsid size, and the presence of contaminants.

• ddPCR (droplet digital PCR) is a highly sensitive method for quantifying the amount of AAV in a sample. It allows accurate and precise measurement of AAV genome copies per milliliter.
• qPCR (quantitative PCR) is a method for detecting and quantifying Mycoplasma contamination in a sample. Mycoplasma is a common contaminant in cell cultures and can affect AAV quality, so confirming its absence is important.
• Capillary Electrophoresis (CE) is a technique used to analyze the integrity of the AAV genome. CE can detect the presence of truncated or rearranged AAV genome fragments, which can affect AAV potency and safety.
• SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and silver stain are methods for analyzing the size and purity of the AAV capsid protein. These methods can detect the presence of impurities and ensure that the capsid size is consistent with the expected size for the AAV serotype.
• TEM (transmission electron microscopy) and AUC (analytical ultracentrifugation) are methods for determining the proportion of empty capsids in the AAV sample. Empty capsids are undesirable as they do not contain the therapeutic payload and can reduce the efficacy of the AAV vector. Monitoring the empty capsid rate is important to ensure the potency and quality of the AAV product.

The combination of these tests allows for a comprehensive assessment of the AAV quality, purity, and safety, ensuring that the AAV produced by PackGene is suitable for use in NHP studies.

Upon receiving the AAV cis plasmid template from our customers, we conduct a double-check of the ITR integrity in addition to verifying the GOI size during clone screening before scaling up the plasmid. After the plasmid preparations are finished, another round of ITR and GOI sequencing is performed to ensure maximum fidelity. You can rest assured that the plasmid we used for NHP grade AAV are of the highest quality and integrity.