Approaches for genetic purity testing of live recombinant viral vaccines using a human adenovirus:rabies model.

Abstract

A two part purity testing regimen for genetically engineered live viral vaccines is described using a human adenovirus 5: rabies glycoprotein gene recombinant as a model vaccine. Initially, restriction endonuclease analysis of the recombinant viral genome verified the integrity of the recombinant construct and identified the vector genome. The second stage employed the polymerase chain reaction to facilitate a more detailed study of the target rabies glycoprotein cassette. The size of the target region was predicted from known nucleic acid sequence information and compared to that obtained after electrophoresis with molecular weight standards. Digestion of the polymerase chain reaction product with a second restriction endonuclease cleaved the target into a number of small fragments. Resolution of the fragments by gel electrophoresis allowed analysis of the target region alone, verifying its identity and integrity.

• Authors: C Lutze-Wallace, T Sapp, S A Nadin-Davis, A Wandeler
• Journal: Canadian journal of veterinary research = Revue canadienne de recherche veterinaire (Can J Vet Res) Vol. 56 Issue 4 Pg. 360-4 (Oct 1992) ISSN: 0830-9000 [Print] Canada
• PMID: 1477804 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Viral
• Oligodeoxyribonucleotides
• Rabies Vaccines
• Vaccines, Synthetic
• Viral Vaccines
• adenovirus 5, rabies glycoprotein recombinant vaccine
• CCCGGG-specific type II deoxyribonucleases
• Deoxyribonucleases, Type II Site-Specific

Topics

• Adenoviruses, Human (immunology)
• Base Sequence
• Cell Line
• DNA, Viral (analysis, chemistry)
• Deoxyribonucleases, Type II Site-Specific
• Electrophoresis, Agar Gel
• Humans
• Molecular Sequence Data
• Oligodeoxyribonucleotides (chemistry)
• Polymerase Chain Reaction
• Rabies Vaccines (genetics, standards)
• Restriction Mapping
• Vaccines, Synthetic (genetics)
• Viral Vaccines (genetics, standards)