Characterization and quality control of recombinant adenovirus vectors for gene therapy.

摘要: Highly purified recombinant adenovirus undergoes routine quality controls for identity, potency and purity prior to its use as a gene therapy vector. Quantitative characterization of infectivity is measurable by the expression DNA binding protein, an early adenoviral in immunofluorescence bioassay on permissive cells determinant. The specific particle count, key indicator, total number intact particles present compared infectious units. Electron microscopic analysis using negative staining gives qualitative biophysical eluted from anion-exchange HPLC. One assessment accomplished via documented presence relative ratios component proteins well potential contaminants reversed-phase HPLC virus followed protein peak identification MALDI-TOF mass spectrometry subsequent data mining. Verification viral genome performed transgene evaluated vitro systems identity. Production lots are also replication-competent human use. For carrying IL-2 transgene, quantitative demonstrated ELISA cytokine cytotoxic T lymphocyte assay following infection cells. Both analyses show good batch reproducibility under test conditions validated methods.