Adenovirus Vector Purification Service

Adenovirus is a very popular gene delivery tool, which has the advantages of simple operation and high safety. Through long-term efforts, Creative Biolabs' scientific and technical team has established a technology platform centered on adenovirus, including vector construction, packaging, detection, and purification. We will provide comprehensive technical support for your research with the most rigorous attitude. No matter what difficulties you have encountered in the relevant research work, you can contact us to get your customized service.

Introduction of Adenovirus Vector

Adenoviruses are a class of non-enveloped viruses that are centered on a linear DNA duplex and are enveloped by a protein envelope. Because it is widely distributed in human body, serotype is diverse and the toxicity of infection is low. It is one of the most commonly used carriers for transmitting genetic material such as DNA to human cells. It also provides molecular tools for gene therapy for many genetic diseases or cancers caused by genetic mutations, which has great research potential and commercial value. Although the expression time of a single transfection of the adenoviral vector is not long, the amount of expressed protein is very high and can reach 30% of the total amount of protein.

Adenovirus vector purification. Figure 1. Adenovirus vector purification.

Adenovirus Vector Purification Service Protocol

  • Recombinant adenoviral vectors will be constructed according to customer requirements. Generally, the popular AdEasyTM system is used to generate a large number of viral particles that are consisted of a shuttle vector carrying a gene of interest and a vector carrying a virus-requiring gene. The correct recombinant adenoviral vector is selected from these viral particles and sequenced to get the correct candidate.
  • The cells are transfected with the recombinant adenoviral vector correctly sequenced in the previous step, cultured for 2-3 weeks, and lysed to obtain a primary virus lysate with a lower titer, which can be stored at -80°C for standby or immediate amplification.
  • Infecting more cells with primary virus lysate to generate more recombinant vectors to achieve higher titers. Generally, cells will be deformed and aggregated after infection. After the infection rate reaches 50%, the virus can be received and a new round of amplification can be started. Each round of amplification can increase the number of viruses by 10-100 times.
  • The recombinant adenoviral vector purification. The recombinant adenoviral vector purification service we provide to our customers can greatly improve the quality of recombinant virus products, allowing them to be used in vivo experiments. The excess cell debris is typically removed by ultracentrifugation using cesium chloride (CsCl) density gradient combined with ultracentrifugation, and the resulting high titer viral vector is stored at -80°C.
  • Virus titer calculation. The titer of the recombinant vector is determined by measuring absorbance, quantitative PCR, and plaque formation assay.

Service

The construction of recombinant adenoviral vectors is a complex and time-consuming process. Creative Biolabs always provides customers with excellent products and services with strict standards and has accumulated a lot of experience in vector construction and purification. If you have any questions or have any difficulties, you can contact us by email or send us an inquiry to find a complete solution.

For research use only. Not intended for any clinical use.