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- Title
Rapid Adeno-Associated Virus Genome Integrity Analysis by Capillary Gel Electrophoresis.
- Authors
Jane Luo; Guttma, Andras
- Abstract
Adeno-associated virus (AAV) is a propitious gene therapy vector to be translated to the clinical practice and pursued accordingly by the biopharmaceutical industry. However, genome integrity and nucleic acid impurities in the final product may represent a problem with the therapeutic use. In this paper, two AAV genome integrity analysis workflows are introduced. The standard sample preparation protocol (~2.5 h) included benzonase treatment to degrade any nucleic acids outside of the AAV capsid, which were then filtered out. The residual AAV capsids were digested by proteinase K, followed by a DNA purification process involving guanidine hydrochloride. The accelerated protocol (~15 min) utilized the DNA purification step only. In both workflows, the released DNA was analyzed by capillary gel electrophoresis with a laser induced fluorescent detection (CGE-LIF) in less than 15 min with = 0.33% RSD on migration time and = 2.06% RSD on peak area reproducibility.
- Subjects
ADENO-associated virus; CAPILLARY electrophoresis; PROTEINASES; NUCLEIC acids; BIOPHARMACEUTICS; GUANIDINIUM chlorides; GENOMES; INTEGRITY
- Publication
LC-GC Europe, 2020, Vol 33, Issue 10, p33
- ISSN
1471-6577
- Publication type
Article