Cat.No. : AAV00270Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 9 Storage: -80 ℃
| Cat. No. | AAV00270Z |
| Description | AAV serotype 9 particles contain firefly luciferase under CAG promoter. |
| Reporter | Luc |
| Serotype | AAV Serotype 9 |
| Application | 1. Determination of optimal MOI (multiplicity of infection), administration methods etc. 2. Detection of the infection efficiency of the AAV serotype against a specific cell type or tissue. 3. Using reporter genes to visualize the distribution and expression of AAV vectors in live animals, helping assess the biodistribution and persistence of gene delivery. |
| Titer | Varies lot by lot, typically ≥1x10^12 GC/mL |
| Size | Varies lot by lot, for example, 30 μL, 50 μL, 100 μL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality AAV particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between AAV particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in the production of AAV, especially for applications in animal studies and gene therapy. Effective endotoxin quality control is essential in the development and manufacturing of AAV particles. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced AAV particles to ensure regulatory compliance. |
| Purity | AAV purity is critical for ensuring the safety and efficacy of AAV-based applications.AAV capsids are composed of three main protein components, known as viral proteins: VP1, VP2, and VP3. These proteins play a critical role in the structure and functionality of the AAV capsid. Monitoring the VP1, VP2, and VP3 content in AAV preparations is essential for quality control in AAV production. Our AAV particles are tested for showing three clear bands of VP1, VP2 VP3 by SDS-PAGE. |
| Sterility | The AAV virus samples are inoculated into the cell culture medium for about 5 days to detect bacterial and fungal growth. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of AAV to deliver genetic material into target cells or tissues, and assess gene expression and functional activities. |
| Empty vs. Full Capsids | Based-on our proprietary AAV production and purification technology, Creative Biogene can always offer AAV particles with high ratio of full capsids. If required, we can also assess the ratio for a specifc lot of AAV particles by transmission electron microscopy (TEM) or other methods. |
Among AAV serotypes, AAV serotype 9 (AAV9) stands out for its ability to effectively target multiple tissues and therefore has great potential for clinical applications. Here, researchers explore a novel purification strategy utilizing Dynabeads™ CaptureSelect™ magnetic beads. AAV9 magnetic beads capture AAV9 with high specificity and recovery rates between 70% and 90%, whereas AAVX magnetic beads do not bind to AAV9. By continuously interacting with AAV in solution, these magnetic beads enhance the clearance of genomic DNA and plasmids even in the absence of endonucleases. The beads can be regenerated at least eight times, and used beads can be stored for up to six months and reused without significant loss in recovery rates. The in vivo potency of AAV9-purified vectors was comparable to that of iodixanol-purified vectors.
To confirm the potency of magnetic affinity bead-purified AAV9, male C57BL/6JInv mice were injected with 5 × 1010 vg/mouse of magnetic affinity bead-purified AAV9-CAG-GFP and AAV9-CAG-Luc via the tail vein. The same dose of the corresponding iodixanol-purified AAV was also used as a positive control. At the endpoint, 9 weeks after AAV injection, livers were harvested and subjected to ex vivo green fluorescent protein (GFP) imaging (Figure 1A) and GFP signal quantification (Figure 1B). The results showed that AAV9-CAG-GFP purified using magnetic affinity beads was functional, and the quantified GFP signal was approximately 1.65-fold lower than that of iodixanol-purified AAV9-CAG-GFP. The gene copy number in the liver of mice injected with magnetic bead-purified AAV9-CAG-GFP was lower than that of iodixanol-purified AAV9-CAG-GFP (Figure 1C).
In addition, the researchers used non-invasive imaging to monitor luciferase signals in AAV-injected mice (Figure 1D). As shown in Figure 1E, luciferase signals were quantified from week 1 to week 9. Unlike the results of GFP imaging, the luciferase signals between magnetic bead-purified AAV9-CAG-Luc and iodixanol-purified AAV9-CAG-Luc showed more similar intensities in vivo. This was further confirmed by quantification of liver AAV genome copy number for AAV9-CAG-Luc vectors, which showed that magnetic affinity bead-purified AAV9-CAG-Luc and iodixanol-purified AAV9-CAG-Luc had similar average values (Figure 1F).
Figure 1. In vivo bioactivity of AAV9-purified by magnetic affinity beads compared to iodixanol-purified AAV9. (Sia K C, et al., 2024)
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