Continuous Capture of recombinant AAV Particles Using Twin-Column CaptureSMB

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Recombinant adeno-associated viruses (rAAVs) have gained increasing importance in gene therapy due to their safe and precise gene delivery. However, certain indications require substantially higher vector doses, pushing manufacturing capacity and cost of goods (COG) to its limits. In this study, we present for the first time a continuous twin-column capture process (CaptureSMB) enabling direct purification of rAAV5 from unprocessed perfusion harvest without prior concentration or processing. This approach differs fundamentally from conventional batch workflows which typically mandate clarification and concentration before affinity capture and offers a novel process integration in viral vector manufacturing. A single-column batch capture process was developed first and subsequently compared to continuous CaptureSMB configurations. Optimized CaptureSMB operation achieved consistent yields over four cycles, with recoveries exceeding batch operation (+ 14.3%) with concomitant higher productivity (+ 11.4%) and reduced buffer consumption (- 79.2%). Critical quality attribute analysis showed lower host cell protein levels and lower residual DNA in early CaptureSMB cycles, while full capsid ratios, thermal stability and transduction efficiency of rAAV5 particles remained unaltered across cycles and process modes. These findings highlight that continuous twin-column CaptureSMB directly from perfusion harvest can not only improve yield and manufacturing efficiency but also maintain and in some respects enhance product quality. This novel strategy provides a promising route to address manufacturing capacity and cost challenges in rAAV gene therapy production.

Authors 11

1. Julia M Mueller

   University of Basel · FHNW University of Applied Sciences and Arts Northwestern Switzerland

   Affiliation as printed

   Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

2. Daniela Tobler

   FHNW University of Applied Sciences and Arts Northwestern Switzerland

   Affiliation as printed

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

3. Jules Buehler

   FHNW University of Applied Sciences and Arts Northwestern Switzerland

   Affiliation as printed

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

4. Damian Hauri
   Affiliation as printed

   ChromaCon AG, Zürich, Switzerland

5. Ralf Plieninger Aachen Institute of Applied Microbiology

   FHNW University of Applied Sciences and Arts Northwestern Switzerland · RWTH Aachen University

   Affiliation as printed

   Institute of Applied Microbiology, RWTH Aachen University, Aachen, Germany

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

6. Sven Göbel

   FHNW University of Applied Sciences and Arts Northwestern Switzerland

   Affiliation as printed

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

7. Efe Saygili

   FHNW University of Applied Sciences and Arts Northwestern Switzerland · Politecnico di Milano

   Affiliation as printed

   Department of Chemistry, Materials and Chemical Engineering “Giulio Natta”, Politecnico di Milano, Milan, Italy

   Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

8. Ryosuke Takahashi

   YMC Co., Ltd. (Japan)

   Affiliation as printed

   YMC CO., LTD., Kyoto, Japan

9. Sebastian Vogg
   Affiliation as printed

   ChromaCon AG, Zürich, Switzerland

10. Thomas Müller‐Späth
    Affiliation as printed

    ChromaCon AG, Zürich, Switzerland

11. Thomas K. Villiger corresponding

    FHNW University of Applied Sciences and Arts Northwestern Switzerland

    Affiliation as printed

    Institute of Pharma Technology and Biotechnology, School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland