Continuous Affinity Purification of Adeno-Associated Virus Using Periodic Counter-Current Chromatography

João P. Mendes, Magnus Bergman, Anita Solbrand, Cristina Peixoto, Manuel J.T. Carrondo, Ricardo J.S. Silva

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)


Replacing batch unit operations of biopharmaceuticals by continuous manufacturing is a maturing concept, with periodic counter-current chromatography (PCC) favoured to replace batch chromatography. Continuous affinity capture of adeno-associated virus (AAV) using PCC has the potential to cope with the high doses required for AAV therapies thanks to its inherent high throughput. The implementation of continuous AAV affinity capture using a four-column PCC process is described herein. First, elution buffer screening was used to optimize virus recovery. Second, breakthrough curves were generated and described using a mechanistic model, which was later used to characterize the loading zone of the PCC. The experimental runs achieved a stable cyclic steady state yielding virus recoveries in line with the optimized batch process (>82%), with almost a three-fold improvement in productivity. The PCC affinity capture process developed here can bolster further improvements to process economics and manufacturing footprint, thereby contributing to the integrated continuous manufacturing concept.

Original languageEnglish
Article number1346
Issue number7
Publication statusPublished - Jul 2022


  • adeno-associated virus
  • affinity purification
  • continuous processing
  • gene therapy
  • periodic counter-current chromatography


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