Human Gene Therapy. 2022 Jan
Yi Pu, Russell Katz, Yunqiu Chen, Vic Kostrubsky, Pete Clarner, Shih-Ching Lo, Zoran Sosic, and Bernice Yeung
Products used in the paper Details Operation
AAV vector packaging Recombinant AAVs were produced by PackGene
(Worcester, MA) or Biogen (Cambridge, MA).
Request Quote

Research Field: New method for AAV safety (in vitro)

Keywords: residual iodixanol, drug safety, adeno-associated virus, purification, mass spectrometry, liquid chromatography


Adeno-associated viruses (AAVs) are nonenveloped viruses that have become popular gene transfer vectors to deliver DNA to target cells in clinical gene therapy. Iodixanol-based density gradient is one of the widely used purification methods for serotype-independent AAVs. However, residual iodixanol in AAV could be a safety concern, and further purification to remove this process-related impurity is typically needed. An analytical assay with high sensitivity is essential for the detection of residual iodixanol to ensure the safety of AAV products. We developed a liquid chromatography-mass spectrometry method with the limit of quantification of 0.01 μg/mL for residual iodixanol measurement in AAVs. The method also demonstrated linearity over four orders of magnitude that allows quantifying a high iodixanol concentration in in-process samples with excellent recovery and accuracy. In addition, we further explored a highly efficient purification method for removal of the residual iodixanol, to minimize the safety concern from iodixanol as a process impurity.

Popular Services

AAV Packaging Service

AAV Analytical Service

Vector Design