Adenovirus is a spherical, non-enveloped virus with a diameter of approximately 70-90nm. Its capsid has a 20-sided icosahedral symmetry and is composed of 252 capsomers, including 240 hexon capsomers and 12 penton capsomers. The fibrous structure protruding from the surface of the capsid is based on penton proteins, with 12 fibers extending from the surface of the capsid. The head regions at the tips of these fibers can bind to receptors on the surface of host cells, playing a crucial role in the virus infection process. Adenovirus replication does not rely on host cell division and has a broad host range. As a professional plasmid manufacturing company, hanbio's adenovirus packaging services are designed to provide high-quality, reliable viral vectors for a variety of applications, including gene therapy, vaccine development, and basic research.
Adenovirus packaging involves cloning the target fragment into an adenovirus expression vector. The expression vector and packaging plasmids are introduced into adenovirus packaging cells for packaging through recombination. There are two main types of recombination for adenovirus packaging: one occurs in bacteria (BJ5183), known as the AdEasyTM adenovirus recombination system, and the other occurs in QBI-293A cells, known as the AdMax adenovirus recombination system. The AdMax system is simple to operate, has high recombination efficiency, produces high virus yields, and enables high expression levels of the target gene.
Construct adenovirus vector and target vector;
Co-transfect HEK293 cells;
Adenovirus collection: Generally, cytopathic effects (CPE) appear 7-10 days post-transfection. When 80% of cells exhibit CPE, collect the supernatant and cells, undergo repeated freeze-thaw cycles, and then centrifuge. After collecting the supernatant, filter it through a membrane and store at -80°C;
Adenovirus amplification: Generally, 80-90% CPE appears 2-3 days post-infection. Collect the cells, undergo repeated freeze-thaw cycles, and then centrifuge. After collecting the supernatant, filter it through a membrane and store at -80°C;
Adenovirus concentration and purification: Use the PEG8000 concentration method;
Identification of recombinant adenovirus: Include tests for virus titer, plasmid residue, host DNA residue, BSA residue, mycoplasma detection, chlamydia detection, endotoxin detection, and bacterial and fungal detection.
When conducting adenovirus packaging experiments, wear lab coats, safety goggles, masks, and gloves. Ensure no skin is exposed and operate within a biological safety cabinet (BL-2 level).
If contamination by the virus occurs inside the biological safety cabinet, immediately clean the area with 70% ethanol and 1% SDS solution.
Dispose of used pipette tips, cell culture dishes, EP tubes, and waste from adenovirus packaging experiments after disinfecting and sterilizing with disinfectants.
After completing adenovirus packaging experiments, thoroughly clean your hands.
