Scalable dual column cation exchange affinity chromatography based platform process for recombinant protein...

Area of Science:

• Biochemistry
• Protein Purification
• Biotechnology

Background:

• Recombinant protein purification is crucial for research and therapeutics.
• Existing methods often face challenges with purity, tag removal, or scalability.
• Novel affinity tags and purification strategies are needed to overcome these limitations.

Purpose of the Study:

• To develop and validate a novel dual-column purification strategy for recombinant proteins.
• To demonstrate the efficacy of a combined heparin-binding tag (HB-tag) and self-removing iCapTag™ system.
• To provide a scalable platform for obtaining highly pure, tagless proteins.

Main Methods:

• A dual-column chromatography process was designed using cation exchange and iCapTag™ split-intein affinity resins.
• Proteins were initially captured using the HB-tag on a cation exchange resin.
• Subsequent purification and tag removal were achieved using the iCapTag™ system with overnight incubation.

Main Results:

• The dual-column method successfully purified diverse proteins including eGFP, βgal, MBP, and βlac.
• High purity levels and effective host cell DNA clearance were achieved.
• The process demonstrated scalability and efficient tag self-removal.

Conclusions:

• The proposed tandem affinity tag system offers a versatile and efficient platform for recombinant protein purification.
• This method combines the benefits of ion exchange capacity and affinity tag selectivity.
• The technology enables the production of tagless proteins suitable for various downstream applications.