Modular high-throughput platform for the purification of scFvs and multispecific antibody-based therapeutics

Automated ÄKTA system-based purification protocols were developed to streamline and accelerate protein production of single-chain variable fragments and multispecific antibodies. The protocols utilize ASX-560 and ALIAS autosampler systems to automate sample injections during overnight and idling times and incorporate one-, two- and three-column schemes to facilitate purification workflows. Additionally, the novel membrane affinity chromatography technology was employed to decrease residence times by increasing flowrates, shortening purification method run time per cycle. As a result, the implementation of versatile and automated multi-column purification protocols enabled the cost-effective and rapid capture of up to 51 single-chain variable fragments per day with high monomeric purities >90 %. Automated affinity chromatography protocols facilitated the daily purification of up to 22 multispecific antibodies. To increase the monomeric target purity of selected single-chain variable fragments and multispecific antibodies, size-exclusion chromatography was automated using the ALIAS Bio PREP autosampler system, enabling the polishing of up to 15 single-chain variable fragments or multispecific antibodies per day.