The Viral Vector Laboratory is dedicated to the production of recombinant viral vectors required for preclinical vaccine studies and other research purposes. One of the key challenges in the use and development of recombinant viral vectors is achieving sufficient yield and quality. To address this, the focus is on producing viral vectors that are both cost-effective and of high quality.
The production process of recombinant viral vectors begins with the creation of a plasmid containing the target gene, using molecular biology techniques. This is followed by a transfection process, where the viral vector capable of producing the target protein is generated. The resulting viral vector is then amplified through serial passages. Once tests for identification and potency are successfully completed, virus stocks are prepared. A standard production process yields purified adenoviral vectors at the appropriate titer, using Ad5-CMV vectors.
The viral vectors produced in this laboratory undergo the following quality control tests to ensure their quality and reliability:
Stability:
Expression of the target protein is confirmed via immunofluorescence analysis, while whole-genome sequencing is used to verify genetic stability.
Potency:
Infectious viral titer is measured using an immunotitration kit. The total viral particle count is determined through spectrophotometry and quantitative PCR (qPCR).
RCA (Replication-Competent Adenovirus) Analysis:
In this analysis, replication-competent adenovirus is assessed using A549 cells, with wild-type Adenovirus type 5 serving as the positive control (Ad5, ATCC VR-5). This test ensures the produced adenoviral vectors are replication-incompetent.
