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Thermo- and pH-Responsible Gels for Efficient Protein Adsorption and Desorption
Izabela Poplewska1, Beata Strachota2, Adam Strachota2
1Department of Chemical and Process Engineering, Rzeszów University of Technology, 35-959 Rzeszów, Poland.
Molecules (Basel, Switzerland)
|October 26, 2024
Summary
This study shows that temperature and pH-sensitive hydrogels can effectively separate proteins. The hydrogels demonstrate tunable protein adsorption and release, offering a salt-free alternative for biomolecule purification.
Area of Science:
- Materials Science
- Biochemistry
- Separation Science
Background:
- Protein adsorption on hydrogels is influenced by hydrogel properties and protein characteristics.
- Thermo- and pH-sensitive hydrogels offer tunable environments for biomolecule interactions.
- Developing efficient and environmentally friendly protein separation methods is crucial.
Purpose of the Study:
- To investigate protein adsorption behavior on poly(N-isopropylacrylamide-co-sodium methacrylate)-based hydrogels.
- To evaluate the influence of temperature and pH on protein adsorption and desorption.
- To explore the potential of these hydrogels for pH-mediated protein separation.
Main Methods:
- Synthesized thermo- and pH-sensitive hydrogels with varying compositions.
- Studied protein adsorption of bovine serum albumin (BSA), ovalbumin (OVA), lysozyme (LYZ), and a monoclonal antibody (mAb2) at different temperatures (5, 20, 37 °C) and pH (4.5, 7, 9.2).
- Analyzed hydrogel swelling, zeta potential, and protein adsorption capacities.
Main Results:
- Hydrogel properties (zeta potential, swelling) were sensitive to temperature, pH, and composition.
- Protein adsorption varied significantly based on protein properties (pI, charge, hydrophobicity) and environmental conditions.
- LYZ showed maximum adsorption at pH 7, BSA/OVA at pH 4.5, and mAb2 at pH 9.2, indicating pH-selective separation.
- Adsorption affinity increased with temperature due to synergistic electrostatic and hydrophobic interactions, especially for mAb2 (20-fold increase).
Conclusions:
- The developed hydrogels enable pH-mediated separation of proteins with different charges under mild, aqueous conditions.
- Temperature increases enhance protein adsorption, while temperature reduction or pH increase facilitates salt-free protein release.
- These hydrogels present a promising platform for selective protein purification and recycling.
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