Intensified AAV purification framework

Personalized medicine is entering a new period with the most recent developments observed in the field of gene therapy. Direct and targeted in vivo delivery of the gene payload using viral vectors is one of the most promising methods and has gained substantial momentum over the last decade with the first approved therapies such as Glybera, Luxturna or Zolgensma.

However, depending on the indication and respective incidence, the estimated annual vector demand can take values between 10E13 vg (Leber’s congenital amaurosis) up to 5E20 vg (Duchenne muscular dystrophy). Manufacturing such amounts, although technically feasible, places substantial pressure in manufacturing processes.

OPPURE tackles the existing gap between scales and manufacturing processes through the following project goals: (i) simplify process architecture, improving productivity and flexibility by using affinity purification; (ii) immobilize affinity ligands in magnetic beads, optimizing ligand density, capacity, and the operating conditions ensuring binding and release of AAV; (iii) intensify purification by capturing AAV in the bioreactor vessel reducing the number of unitary operations and (iv) develop a polishing process for improved product purity.

The main activities of the proposed project can be divided into three groups: (i) Affinity capture performed directly in the bioreactor, supported by the use of Superparamagnetic beads; (ii) Rapid cycling chromatography for polishing using highly permeable convective chromatographic media delivering throughputs similar to those observed in Periodic counter-current chromatography; and (iii) A rigorous modeling framework for increased process understanding and to facilitate process scaling.

In summary, OPPURE is an intensified purification framework that can cope with the different demand levels and manufacturing scales. By doing so, it is possible to target not only a reduction in the cost of goods but also set the ground for faster development of these therapies.