Optimizing Adeno-Associated Virus (AAV) Capsid Protein Analysis Using UPLC and UPLC-MS

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Ximo Zhang, Stephan Koza, Ying Qing Yu, Weibin Chen
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Application Notes
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Application Notes
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Cell and Gene Therapy

In this work, an optimized RPLC method was developed using DFA as a mobile phase modifier for improved separation of AAV capsid proteins, while retaining adequate mass spectrometry sensitivity. The combination of the mobile phases and judicious selection of column chemistry results in a chromatographic separation that facilitates intact mass measurement of individual VP isoforms and their variants. The developed method also renders a means to quantify VP stoichiometry through LC-optical response at sub-microgram levels. Overall, results acquired on the BioAccord System show that this platform can deliver the structural information necessary to improve the understanding of gene therapy products throughout product development and commercialization.

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