Journal List > J Bacteriol Virol > v.38(4) > 1033901

Park, Sohn, and Paik: Production and Quality Control of Adenoviral Vectors for Clinical Trials


The importance of recombinant adenoviral vectors for the development of gene therapy and prophylactic and therapeutic vaccines has led to efforts for process development of large scale production of clinically safe adenoviral vectors. First of all, cell lines producing replication incompetent adenoviral vectors required for clinical application have been developed and the concept of banking and characterization of cell lines and adenoviral vectors has been established. In order to meet the need of amount of adenoviral vectors for clinical trials, various large scale suspension culture methods using serum-free media have been developed along with development of large scale purification methods using chromatography instead of cesium chloride method. In addition, methods for the quality control of adenoviral vectors have been established and applied for the clinical lots.


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Figure 1.
Adenovirus packaging cell line and adenoviral vector. A) HEK293 cell: Appearance of RCA (replication competent adenovirus) by homology (gray box) between packaging cell line and vector. B) PER.C6 cell: No appearance of RCA due to the absence of homology between packaging cell line and vector.
Table 1.
Analysis Items of Cell Line Bank and Virus Bank
Analysis items Cell line bank Virus bank Note
Morphology + + +  
Identity - isoenzymes + +  
DNA fingerprinting + +  
Karyology (diploid cell lines) +    
Bacterial and fungal contamination + + + +  
Mycoplasma + + + + +  
In vitro virus assay (CPE, HA) + + + + CPE: viral cytopathic effects HA: hemadsorption
In vivo virus assay + +  
Electronmicroscopy +  
Specific viruses +  
Bovine viruses + + + If bovine serum has been added.
Porcine viruses + + + If porcine trypsin has been added.
Retroviruses (PCR-based RTase) + +  
Endotoxins (LAL)  
Host cell DNA  
Contaminating proteins  
Virus titration + +  
Table 2.
Category and Standards for Quality Evaluation of Adenoviral Vector
Test items   Standard
Appearance   Self-standard
pH   Self-standard
Sterility test   No Sterile
Endotoxin test   Self-standard (commonly <5 EU/ml)
  Virus titer (OD260) Self-standard
Content Virus titer (HPLC) Self-standard
  Infectious titer Self-standard
Verification Restriction enzyme map Expected band pattern
  Host DNA test Self-standard
Purity test Host protein test Self-standard
  Aggregation (A320/A260) Self-standard
Potency test Infectious titer/virus particle <30 (USA FDA standard)
  BSA content Self-standard
Residual materials during manufacturing process Benzonase content Self-standard
  Triton X-100 content Self-standard
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