Development and Application of a Liquid Chromatography-Mass Spectrometry Method for Residual Iodixanol Quantification in AAV-Based Gene Therapy Product Development.

Autor:	Pu Y; Analytical Development, Cambridge, Massachusetts, USA., Katz R; Gene Therapy-Process Development, Cambridge, Massachusetts, USA., Chen Y; Analytical Development, Cambridge, Massachusetts, USA., Kostrubsky V; Preclinical Safety, Cambridge, Massachusetts, USA., Clarner P; Gene Therapy Accelerator Unit, Cambridge, Massachusetts, USA., Lo SC; Research, Biogen, Inc., Cambridge, Massachusetts, USA., Sosic Z; Analytical Development, Cambridge, Massachusetts, USA., Yeung B; Analytical Development, Cambridge, Massachusetts, USA.
Jazyk:	angličtina
Zdroj:	Human gene therapy [Hum Gene Ther] 2022 Jan; Vol. 33 (1-2), pp. 103-108. Date of Electronic Publication: 2021 Sep 20.
DOI:	10.1089/hum.2021.136
Abstrakt:	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.
Databáze:	MEDLINE
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