Metabolic Biotinylation and Purification of Multiple AAV Vector Serotypes

Adeno-associated virus (AAV), also known as adeno-associated virus, belongs to the family Parvoviridae and the genus Dependovirus. Adeno-associated viruses are simple, single-stranded DNA deficient viruses that require a helper virus to replicate. Adeno-associated virus can infect a variety of cells, showing great application potential in human gene therapy. AAV 2 is currently the most widely used virus for gene delivery. Biotin is a component of the microbial B₂ complex, a cofactor for fatty acid and nucleic acid biosynthesis. Biotin is necessary for the metabolic process of biological problems. Biotin is covalently assembled to the biotin carrier protein through the action of biotin ligase, a biotin ligase holocarboxylase synthetase (HCS) produced by the BirA gene in Escherichia coli or in mammalian cells. Enzymes encoded by genes. Biotin has a very high affinity for avidin, a tetrameric glycoprotein secreted in egg white, which can be used for cross-linking, staining or targeted therapy. The chemical process of biotinylation often results in non-specific labeling and inactivation of the target protein. Biotinylation is performed in vitro or in vitro by the enzymatic action of biotin ligase.

The Services

Alfa Chemistry has a professional team that has long been engaged in biotinylation-related research and can provide efficient biotinylation reactions, including but not limited to:

• Affinity purification of immobilized monomeric avidin by biotinylated AAV
• Biological activity assays of biotinylated AAV vectors
• Construction of modified AAV helper plasmids
• Western blot detection
• Solid-phase binding of biotinylated AAV to avidin

Biotin-avidin system

The biotin-avidin system can carry out multi-level amplification effect, and can carry out efficient immunodiagnosis, biomarker, quantitative detection and localization observation. Biotin is widely distributed in animal and plant tissues. The structure of biotin molecule contains a ring structure, which can combine with avidin. After chemical modification, biotin can be called an active group for efficient biological applications.

Adeno-associated virus and gene therapy

At present, tumor is still a major disease that endangers human health, and efficient treatment for it is of great significance for maintaining human health. Gene therapy is the introduction of foreign genes into target cells, which can improve diseases caused by gene defects. Gene therapy has expanded from single-gene genetic diseases to malignant tumors, infectious diseases, cardiovascular diseases, autoimmune diseases, metabolic diseases and other major diseases. Adeno-associated virus is the carrier of gene therapy, and various methods have been developed for efficient gene therapy.

How to choose an adeno-associated virus vector?

Adeno-associated virus (AAV) belongs to the genus Dependovirus of the Parvoviridae family, a single-stranded DNA virus, and is the smallest and simplest animal virus. The following factors need to be considered in selecting the appropriate adeno-associated virus vector:

• Choose the right promoter
• Design payload

What are the limitations of adeno-associated virus?

• Low expression level in vitro experiments
• Takes longer to express
• Carrier capacity is small

At Alfa Chemistry, researchers are committed to providing customers with high-quality services for efficient biotinylation in vivo and in vitro. If you are interested in our services, please contact us immediately.

Reference:

1. Gregory. S. A.; et al. Metabolic biotinylation provides a unique platform for the purification and targeting of multiple AAV vector serotypes. MOLECULAR THERAPY. 2006, 14(1): 97-106.

※ It should be noted that our service is only used for research, not for clinical use.