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Peptide affinity reagents for AAV capsid recognition and purification

Gene Therapy volume 18, pages 1020–1024 (2011)Cite this article

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Abstract

We report the discovery of AAV capsid-binding peptides identified through phage panning. The heptapeptide motif GYVSRHP selectively recognized AAV serotype 8 capsids and blocked transduction in vitro. Recombinant AAV8 vectors were purified directly from crude cell lysate and supernatant through sequential application of peptide affinity and anion exchange chromatography. Peptide affinity reagents may serve as useful alternatives to monoclonal antibodies in AAV capsid recognition, and offer readily scalable solutions for purification of clinical grade AAV vectors.

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Acknowledgements

We would like to thank Stephen Soltys at the UNC vector core for assistance with column chromatography and Swati Yadav for qPCR analysis. This study was supported by grants from the American Heart Association, NIH (HL089221) and ARRA funds (HL089221-S1/S2) awarded to AA.

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Authors and Affiliations

  1. Gene Therapy Center, The University of North Carolina, Chapel Hill, NC, USA

    N Pulicherla & A Asokan

  2. Department of Genetics, The University of North Carolina, Chapel Hill, NC, USA

    A Asokan

  3. Molecular and Cellular Biophysics Program, The University of North Carolina, Chapel Hill, NC, USA

    A Asokan

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  1. N Pulicherla
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  2. A Asokan
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Correspondence to A Asokan.

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The authors declare no conflict of interest.

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Supplementary Information accompanies the paper on Gene Therapy website

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Pulicherla, N., Asokan, A. Peptide affinity reagents for AAV capsid recognition and purification. Gene Ther 18, 1020–1024 (2011). https://doi.org/10.1038/gt.2011.46

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