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

Area of Science:

• Biochemistry
• Protein Engineering
• Chromatography

Background:

• Protein production requires efficient purification methods for biologics like single-chain variable fragments (scFv) and multispecific antibodies.
• Current purification workflows can be time-consuming and labor-intensive, hindering rapid development.

Purpose of the Study:

• To develop and implement automated ÄKTA system-based purification protocols.
• To streamline and accelerate the production of scFv and multispecific antibodies.
• To enhance purification efficiency and purity using novel chromatography techniques.

Main Methods:

• Development of automated ÄKTA system protocols utilizing ASX-560 and ALIAS autosampler systems.
• Implementation of one-, two-, and three-column purification schemes.
• Application of membrane affinity chromatography to reduce residence times and increase flow rates.
• Automation of size-exclusion chromatography for polishing using the ALIAS Bio PREP autosampler.

Main Results:

• Achieved cost-effective and rapid capture of up to 51 scFv per day with >90% monomeric purity.
• Facilitated daily purification of up to 22 multispecific antibodies using automated affinity chromatography.
• Enabled polishing of up to 15 scFv or multispecific antibodies per day with enhanced purity.

Conclusions:

• Automated multi-column purification protocols significantly improve the speed and efficiency of scFv and multispecific antibody production.
• Novel membrane affinity and automated size-exclusion chromatography enhance purification yields and purity.
• These advancements support cost-effective and rapid biologics development.