We offer the production of adenovirus, its amplification and purification for the wild type adenovirus 5 or, the virus that the client wants to produce and also the possibility of creating genetic engineering, the virus from the gene of interest of the client. Based on the initial stock of wild type Ad5 obtained from the American Type Culture Collection (ATCC), HEK293 cells are infected to produce the initial seed that will infect the stock to be produced. The method of generation of recombinant Adenoviruses is by homologous recombination in bacteria and subsequent expansion and purification by double gradient of Cesium Chloride followed by a Sephadex G-25 column. Delivery volumes are limited and depend on the virus titer. The estimated delivery time of the service, depending on the complexity, is as follows:
- 20 days amplification and purification
- 60 days a complete construction
Equipment:
Ultracentrifuge L7-65 Beckman 27493M08 - Class II biological safety cabinets
Applications
The advance in the knowledge of the virus allowed progress in the development of viruses or viral vectors as transport tools for genes of therapeutic interest for gene therapy and protein expression, in the field of human and animal health. Among the most used viral vectors, adenoviruses are found. The applications of viral vectors for gene therapy are used for the treatment of genetic diseases and also for acquired diseases, mainly in cancer and AIDS, and to a lesser extent, for the treatment of cystic fibrosis and metabolic diseases.
Adenoviruses may act as reporters expressing proteins that, at a given wavelength, emit colors such as green, red, blue which can be observed by microscopy. Moreover, they may carry genes that express proteins which are abnormal in a cell and may regenerate the lost biological function. In particular, adenovirus models may be used in the development of genetic therapies, drug testing, correction of abnormal metabolic pathways, gene silencing, tumor lysis in research phases.
Advantages
Viral vectors are the most used, due to the efficiency of gene transfer compared to non-viral vectors. Adenoviruses have the following advantages and specificities:
- Great capacity to incorporate large genes
- They can infect cells in division or not in division
- Ability to produce high titers of recombinant virus
- Significant immune response
- In vivo transient gene expression
