AAV Production Systems
Four production scales built on a unified triple plasmid transfection platform — from early vector validation to IND-enabling clinical manufacture, with consistent QC at every tier.
All tiers ship in PBS + 5% sorbitol, pH 7.4 (standard) or customer-specified formulation buffer · Storage: −80 °C recommended
Tissue Targeting Guide
Capsid serotype determines tissue tropism, transduction efficiency, and immunogenic profile. Match your target tissue to the optimal AAV variant using the reference guide below.
| Serotype | Primary Tissue | Route | Efficiency |
|---|---|---|---|
| AAV1 | Skeletal muscle, heart | IM, IV |
|
| AAV2 | Retina, liver, brain | Sub-retinal, IC |
|
| AAV5 | Lung, brain (striatum) | Intranasal, IC |
|
| AAV8 | Liver (hepatocyte) | IV, portal vein |
|
| AAV9 | Heart, CNS, muscle | IV, intrathecal |
|
| PHP.eB | CNS (neurons, astrocytes) | IV (mouse only) |
|
| AAVrh10 | CNS, liver, lung | IV, IC, intrathecal |
|
| AAV-DJ | Liver, muscle (in vitro) | IV, IM |
|
Efficiency ratings reflect relative in vivo transduction efficiency via indicated route in rodent models. PHP.eB CNS efficiency is specific to C57BL/6 and BALB/c mice; not validated for NHP. IV = intravenous; IM = intramuscular; IC = intracranial.
Quality Control Assays
Every production lot is characterized using orthogonal analytical methods. CoA reports are issued with all deliverables.
Frequently Asked Questions
Technical questions about AAV production, QC, and project logistics. Can't find what you need? Our PhD scientists respond within one business day.
The AAV genome packaging limit is < 4.7 kb between the two ITRs. This includes your promoter, transgene coding sequence, 5′/3′ UTRs, poly-A signal, and any regulatory elements.
Oversized constructs (> 5.0 kb) result in dramatically reduced titers and a high proportion of truncated genomes. For large genes, consider split-intein approaches (two complementary half-vectors) or miniaturized promoters (e.g., CMV enhancer/CBA, miniCMV, or synapsin variants) to stay within the packaging limit.
Dual-vector trans-splicing strategies can accommodate transgenes up to ~8–9 kb but require co-transduction of both vectors and result in lower overall expression efficiency compared to single-vector approaches.
Serotype selection depends on four factors: target tissue, delivery route, species, and pre-existing immunity in the model.
As a starting point: AAV8 or AAV-DJ for liver; PHP.eB for CNS in C57BL/6 mice (IV); AAV9 for cardiac or broad CNS; AAV1 for skeletal muscle; AAV2 for retinal subretinal injection. For NHP or human cells, avoid PHP.eB — use AAVrh10, AAV9, or AAV5 instead.
If you're unsure, our scientists can recommend a serotype based on your target. Use the Tissue Targeting Guide above or contact our team.
Genome titer (VG/mL) quantifies total viral genomes by ddPCR or qPCR. It includes both full (genome-containing) and empty capsids, and represents the physical particle count regardless of infectivity.
Functional titer (IU/mL or TU/mL) measures transducing units — the number of particles capable of infecting a cell and expressing the transgene. This is determined by in vitro transduction assay on HEK293 or HeLa cells, followed by FACS or qPCR for transgene copies.
For in vivo dosing, genome titer is the standard unit. For in vitro applications and lentiviral comparisons, functional titer is more relevant. A typical VG:IU ratio for well-purified AAV is 10–100:1. Functional titer is available as an add-on assay.
Standard formulation is PBS (pH 7.4) + 5% sorbitol, which provides cryoprotection and maintains capsid integrity during freeze-thaw cycles. Vectors are aliquoted to minimize freeze-thaw cycles and shipped on dry ice.
Custom formulation buffers are available upon request (e.g., Lactated Ringer's for in vivo CNS applications, or BSS for ocular delivery). There is no additional charge for standard alternative buffers; specialized buffer formulation for GMP lots should be specified at project initiation.
Recommended storage: −80 °C for long-term. Stable for up to 12 months at −80 °C with ≤ 3 freeze-thaw cycles. Avoid −20 °C storage.
After project confirmation, you ship your transfer plasmid (ITR-GOI-ITR) as a verified, endotoxin-free maxi-prep at ≥ 1 µg/µL in TE or water. We verify sequence and ITR integrity before initiating production.
If you prefer, we can synthesize and clone your construct directly. Provide your transgene sequence (FASTA or GenBank), preferred promoter, and any required regulatory elements. Custom vector design adds approximately 1–2 weeks to the project timeline.
We require: plasmid sequence map (SnapGene or equivalent), annotated ITR positions, and confirmation that the construct is ≤ 4.7 kb between ITRs.
Yes. Customer-supplied Rep/Cap plasmids encoding novel or engineered capsids (e.g., directed evolution variants, rational design mutants, chimeric capsids) can be used in the triple plasmid system, provided the capsid sequence is compatible with the standard HEK293T production platform.
Note that novel capsids may require capsid-specific process optimization (e.g., adjusted transfection ratios, modified harvest timing) and may yield lower titers than validated serotypes on the first production run. We recommend a pilot-scale feasibility run before committing to large-scale production.
All IP in the resulting vector and remaining cis plasmid belongs to the customer.
Each CoA includes: lot number, production date, serotype, transgene identity, vector genome sequence confirmation, ddPCR titer with raw data, SDS-PAGE gel image, empty/full ratio (Tier 2+), endotoxin value (LAL; Tier 2+), mycoplasma result (Tier 2+), formulation buffer, volume delivered, and pass/fail determination against specification table.
GMP lots additionally include batch manufacturing records, raw material certificates, environmental monitoring data, and full ICH Q6B release results.
Ready to start your AAV project?
Discuss your vector design, production scale, and timeline with our PhD scientists. We'll follow up within one business day with a detailed quote and feasibility assessment.