Establishment of a platform imaged capillary isoelectric focusing (icIEF) characterization method for adeno-associated virus (AAV) capsid proteins
Jiaqi Wu, Chris Heger
Abstract
Herein we describe a platform method to study the charge heterogeneity of adeno-associated virus (AAV) capsid proteins . The method, based on imaged capillary isoelectric focusing (icIEF) with native fluorescence imaging detection, was validated, and can be used as an identity assay and stability indication assay. The protocol was also evaluated following the White Analytical Chemistry principle, its score is 89.7. The method is shown to be platform, with eight different AAV serotypes successfully analyzed. The resulting AAV serotype ‘fingerprints’ can be used for AAV identity and stability indication.
Topics & Concepts
CapsidAdeno-associated virusIsoelectric focusingVirusVirologyCapillary actionCharacterization (materials science)Capillary electrophoresisChemistryBiologyMolecular biologyPhysicsMaterials scienceNanotechnologyBiochemistryGeneVector (molecular biology)Recombinant DNAEnzymeThermodynamicsVirus-based gene therapy researchViral gastroenteritis research and epidemiologyBiosensors and Analytical Detection