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Affinity membrane adsorbers for binding arginine-rich proteins
Heather C S Chenette1, James M Welsh1, Scott M Husson1
1Department of Chemical and Biomolecular Engineering and Center for Advanced Engineering Fibers and Films, Clemson University, Clemson, SC 29634, USA.
Separation Science and Technology
|October 13, 2023
Summary
Researchers developed a new affinity membrane for purifying arginine-rich fusion proteins, a key step for delivering protein cancer therapies. The membrane showed promising protein binding capacity, though further optimization is needed for practical applications.
Area of Science:
- Biotechnology
- Materials Science
- Chemical Engineering
Background:
- Delivering protein-based chemotherapeutics into cancer cells remains a significant challenge in oncology.
- Arginine-rich cell-penetrating peptides offer a potential strategy to enhance intracellular delivery of therapeutic proteins.
- Efficient purification of these engineered fusion proteins is crucial for their development and clinical application.
Purpose of the Study:
- To develop an affinity membrane for the capture chromatography-based purification of arginine-rich fusion proteins.
- To functionalize macroporous membranes with diethyl-4-aminobenzyl phosphonate (D4ABP) ligands for protein binding.
Main Methods:
- Grafting polymers functionalized with diethyl-4-aminobenzyl phosphonate (D4ABP) ligands onto macroporous membrane supports.
- Characterization of D4ABP incorporation using infrared spectroscopy and energy dispersive spectroscopy.
- Measurement of protein binding capacity using lysozyme as a model protein.
Main Results:
- Successful preparation of functionalized membranes bearing D4ABP ligands.
- Demonstrated protein binding capacity of 3 mg lysozyme/mL.
- Indicated the need for further studies on binding kinetics and Arg-selectivity, and for increased binding capacity.
Conclusions:
- The developed affinity membrane shows potential for purifying arginine-rich fusion proteins.
- Further optimization is required to enhance protein binding capacity and selectivity for practical therapeutic applications.
- This approach could facilitate the development of protein-based cancer therapies.
