Cat.No. : AAV00299Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV serotype PHP.B Storage: -80 ℃
| Cat. No. | AAV00299Z |
| Description | AAV serotype PHP.B particles contain GFP under CAG promoter. |
| Reporter | GFP |
| Serotype | AAV serotype PHP.B |
| 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. |
Currently, gene transfer is typically achieved through intraparenchymal viral injections, but these injections target limited brain regions. Here, researchers demonstrate that intravenous delivery of adeno-associated virus (AAV)-PHP.B viral particles penetrates and spreads throughout the neural parenchyma, targeting the central and peripheral nervous systems in a holistic pattern. The researchers then established multiple viral transduction procedures to control gene expression or simply inactivate gene function in the adult nervous system and assessed potential behavioral effects. Based on these results, an effective gene therapy strategy was established to counteract the widespread accumulation of α-synuclein deposits throughout the forebrain in a mouse model of synucleinopathies. Transduction of A53T-SCNA transgenic mice with AAV-PHP.B-GBA1 restored physiological levels of the enzyme, reduced α-synuclein pathology, and produced significant behavioral recovery. Furthermore, studies have shown that AAV-PHP.B brain penetration does not result in evident dysfunctions in blood-brain barrier integrity or permeability. Taken together, the AAV-PHP.B viral platform enables non-invasive, extensive and durable global neural expression of therapeutic genes such as GBA1, providing an invaluable approach to treat neurodegenerative diseases with diffuse brain pathology such as synucleinopathies.
Here, the researchers analyzed blood-brain barrier (BBB) permeability and inflammation after in vivo transduction of AAV-PHP.B. To this end, mice were intravenously injected with a fluorescently conjugated cadaverine dye, a small BBB permeability marker, and the virus AAV-PHP.B-GFP (Figure 1A). Staining of the viral capsid with an AAV-VP3-specific antibody (B1) confirmed that the viral particles were located within the brain endothelium 24 hours after viral delivery (Figure 1B). However, the transduced brain tissue did not show any obvious diffusion of the cadaverine dye (Figure 1C). In addition, GFAP staining did not reveal signs of astrogliosis in the target tissue 2 days after viral transduction (Figure 1C). As a positive control, diffuse cadaverine staining and astrocyte activation were detected in the brain parenchyma of mice treated with kainic acid, which developed seizure-induced BBB permeability and severe inflammation (Figures 1E-1G). These results indicate that AAV-PHP.B brain transduction does not affect BBB permeability in vivo.
Figure 1. AAV-PHP.B brain transduction does not affect BBB permeability in vivo. (Morabito G, et al., 2017)
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AAV-PHP.B's ability to efficiently transduce the central nervous system (CNS) after intravenous injection is a game-changer for our research. It surpassed AAV9 in CNS transduction in specific mouse strains.
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