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Updated: Jan 26, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

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Published on: January 16, 2012

Column-Free Purification Methods for Recombinant Proteins Using Self-Cleaving Aggregating Tags.

Yamin Fan¹, Jackelyn M Miozzi², Samuel D Stimple³

¹William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA. fan.457@osu.edu.

|April 11, 2019

Summary

We developed novel non-chromatographic protein purification methods using self-cleaving elastin-like polypeptide (ELP) or β-roll tags (BRT17) with split inteins. These methods offer scalable, cost-effective purification with high yields for recombinant proteins.

Keywords:

aggregating tags elastin-like polypeptide (ELP)non-chromatographic protein purification self-cleaving tag split intein β-roll tag (BRT17)

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Area of Science:

Biotechnology
Protein Purification
Molecular Biology

Background:

Conventional chromatography for recombinant protein purification is costly and slow.
Current tag removal methods are expensive and not scalable.
Non-chromatographic methods using aggregating tags offer cost reduction potential.

Purpose of the Study:

To develop non-chromatographic purification strategies for recombinant proteins.
To enable efficient and scalable tag removal using engineered split inteins.
To improve protein yields compared to existing methods.

Main Methods:

Utilized elastin-like polypeptide (ELP) and β-roll tag (BRT17) as aggregating tags.
Integrated engineered split inteins for controlled, mild tag cleavage post-purification.
Applied the system to purify β-lactamase (β-lac), sfGFP, streptokinase (SK), and MBP.

Main Results:

Achieved efficient purification of multiple recombinant proteins.
Obtained yields typically ranging from 200-300 µg/mL of cell culture.
Reported overall recoveries between 10-85%, dependent on the target protein.

Conclusions:

Developed a scalable, cost-effective non-chromatographic purification system.
The split intein system prevents premature cleavage and allows controlled tag removal.
The novel strategies enhance protein yields for biotechnological applications.