Transforming mAb purification

Monoclonal antibodies are some of the best-selling biopharmaceuticals, with more than 90 mAbs approved by global regulatory agencies as of December 2019. A prediction from GlobalData indicated that the mAbs market would reach a value of $156 billion in 2020, growing at a CAGR of 10% between 2019 and 2025.¹

So, it is no wonder that there are plenty of options now available when it comes to the capture step using antibody affinity resins. Early Protein A chromatography resins have proven to be efficient, reliable, and easy to implement at any scale. They have a long track record in the industry and are well accepted by regulatory authorities. However, Protein A resins have historically been more sensitive to harsh cleaning conditions than other chromatographic techniques, due to the nature of the protein ligand. This imposes challenges and risk of microorganism contaminations, as the first capture step is exposed to feed with a lot of nutrients and, at the same time, is sensitive to high concentrations of sodium hydroxide (a common cleaning solution). To work around this issue later-generation Protein A resins have become more alkaline stable, allowing for better bioburden control.

At Cytiva we have several Protein A chromatography options to choose from: traditional resin chromatography technologies and the new fibre-based chromatography technology called Fibro PrismA™.

With Protein A chromatography resin, target molecules carried in the bulk feed media must go through two stages of mass transfer before reaching the functional binding surface buried deep within the pores of a chromatography bead. The initial film diffusion, followed by diffusion into the porous structure, results in a flow-rate-limited operation, which presents a fundamental barrier to improving purification productivity.

Instead, the structure of the fibres in the Fibro technology is highly open to facilitate mass transfer without diffusion, like membrane and monolith technologies, but it also yields a high surface area. This is a result of its manufacturing method, an additive process known as electrospinning. These unique features of Fibro allow it to obtain binding capacities of more than 30 mg/mL in seconds of residence time rather than minutes as with conventional bead chromatography (Fig 1).

Fig 1. Fibre chromatography benefits include increased productivity (up to 400g/h/L), up to 50 times increased throughput for research and process development applications.

The Fibro chromatography technology enables rapid cycling antibody purification. In research and process development, this means Fibro PrismA can support up to 20 times increased throughput compared with traditional resins. All this can be done whilst still using the same buffer components and volumes, chromatography systems, infrastructure and ligands as Cytiva’s existing resin chromatography, ensuring a good fit with existing facilities. For example, once a HiTrap™ Fibro PrismA unit is connected correctly to your existing ÄKTA™ system, a full purification cycle with chromatogram can be achieved in less than 5 minutes.

This method speed has not been possible until now and it can really be a game changer for research. LifeArc in the United Kingdom, an early adopter of Fibro PrismA technology, reduced the time to purify mAbs for a COVID-19 project from 14 to 5 days.

To read more on how Fibro PrismA helped LifeArc, please click here.

¹GlobalData (2020) Drugs Database

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