E. coli-produced nanobody fusions as a pan-serotype platform for AAV purification, quantification, and detection

Nanobody AAVX fusion proteins expressed in Escherichia coli enable one-step IMAC purification yielding mostly >90% pure protein and efficient capture of intact AAV2 and AAV5 capsids.

• Why it matters: rAAV research and bioprocessing need versatile, rapid, and cost-effective tools to purify, measure, and detect AAV across serotypes, with adaptable reagents.
• What they did: The team cytosolically expressed E. coli nanobody AAVX with various tags and fusion partners, then used IMAC and agarose crosslinking to purify intact AAV2 and AAV5, quantified by qPCR and ELISA.
• The result: AAVX functioned as an ELISA quantification reagent and, when fused to luciferase, as a detection reagent, and it supported label-free capsid quantification by BLI and fluorescence microscopy detection in fixed HEK-293 cells.

AI summary of the abstract · may contain errors · not medical advice · read the paper.