Sample interview questions: Can you explain the principles of viral vector production and purification for gene therapy applications?
Sample answer:
-
Viral vector production:
- Select an appropriate viral vector system, commonly adeno-associated viruses (AAVs), lentiviruses, or retroviruses.
- Generate recombinant viral vectors using genetic engineering techniques to incorporate the gene of interest.
- Transfect or infect producer cells with the recombinant viral vector DNA or RNA to initiate viral replication and packaging.
- Harvest viral particles from the producer cell culture medium or lysates.
-
Viral vector purification:
- Clarify the viral suspension by centrifugation or filtration to remove cell debris and large particles.
- Concentrate the viral particles using ultracentrifugation, precipitation, or tangential flow filtration.
- Perform size-exclusion chromatography or iodixanol gradient ultracentrifugation to separate viral particles based on size and density.
- Further purification steps may include anion-exchange or affinity chromatography to remove impurities and… Read full answer
Source: https://hireabo.com/job/2_3_34/Tissue%20Culture%20Specialist