Work

IROP: Ionic regulation of permeation for high-yield AAV purification

Year: 2026

Type: article

Abstract: Traditional filtration methods for separating host cell proteins and DNA from adeno-associated viral capsids often result in substantial yield losses. Here, we introduce a new high-yield purification strategy termed Ionic Regulation of Permeation (IROP) for removing major impurities within a single unit operation. Rather than relying on static size exclusion or binding interactions, IROP dynamically modulates AAV transport within a nanoporous membrane. At low ionic strength, AAV permeation was negligible, while high ionic strength markedly increased the sieving coefficient, revealing an electrostatic gating mechanism. AAVs retained under low-salt conditions could not be efficiently recovered by a static high-salt wash, indicating steric-electrostatic trapping within membrane pores. In contrast, high-salt filtration enabled high recovery. Leveraging this mechanism, a two-step IROP process, involving low-salt impurity clearance followed by high-salt AAV recovery, achieved greater than 90% AAV recovery with a greater than 95% reduction in HCPs and host cell DNA.

Source: Journal of Membrane Science Letters

DOI: 10.1016/j.memlet.2026.100120

Authors: Deepraj Sarmah, Scott M. Husson

Institution: Clemson University

Language: English

Topic: Protein purification and stability

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,310

Funders: National Science Foundation, Clemson University