Comprehensive Characterization and Quantification of Adeno Associated Vectors by Size Exclusion Chromatography and Multi Angle Light Scattering

Overview

Journal Sci Rep

Specialty Science

Date 2021 Feb 5

PMID 33542328

Citations 40

Authors

Nicole L McIntosh

Geoffrey Y Berguig

Omair A Karim

Christa L Cortesio

Rolando De Angelis

Ayesha A Khan

Daniel Gold

John A Maga

Vikas S Bhat

Affiliations

Soon will be listed here.

Abstract

Adeno associated virus (AAV) capsids are a leading modality for in vivo gene delivery. Complete and precise characterization of capsid particles, including capsid and vector genome concentration, is necessary to safely and efficaciously dose patients. Size exclusion chromatography (SEC) coupled to multiangle light scattering (MALS) offers a straightforward approach to comprehensively characterize AAV capsids. The current study demonstrates that this method provides detailed AAV characterization information, including but not limited to aggregation profile, size-distribution, capsid content, capsid molar mass, encapsidated DNA molar mass, and total capsid and vector genome titer. Currently, multiple techniques are required to generate this information, with varying accuracy and precision. In the current study, a new series of equations for SEC-MALS are used in tandem with intrinsic properties of the capsids and encapsidated DNA to quantify multiple physical AAV attributes in one 20-min run with minimal sample manipulation, high accuracy, and high precision. These novel applications designate this well-established method as a powerful tool for product development and process analytics in future gene therapy programs.

Citing Articles

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Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus.

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