Optimization of the Preparation of Hydrophilic Poly(DHPMA-co-MBA) Monolithic Capillary Columns: A New Support for Affinity Chromatography
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Buffers
2.2. Monolithic Capillary Column Synthesis
2.2.1. In-Capillary Poly(GMA-co-MBA) Monolith Synthesis
2.2.2. In-Capillary Poly(DHPMA-co-MBA) Monolith Synthesis
2.3. Column Biofunctionalization
2.3.1. Preparation of Streptavidin-Functionalized Monolithic Capillary Columns
2.3.2. Preparation of Concanavalin a Monolithic Capillary Columns
2.4. Nano-LC Experiments
2.4.1. Nano-FAC Experiments: Evaluation of Non-Specific Interactions
2.4.2. Quantification of the Number of Protein Active Binding Sites (Streptavidin or Concanavalin Columns)
3. Results and Discussion
3.1. Synthesis and Characterization of DHPMA-co-MBA Monoliths
3.2. Characterization of the Number of Active Binding Sites after Protein Immobilization
3.3. Evaluation of the Non-Specific Interactions on Poly(DHPMA-co-MBA) Monoliths and Comparison with Poly(GMA-co-MBA) Monoliths
3.4. Separation of Concanavalin Ligands by Weak Affinity Chromatography
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Weight (mg) | % | |
---|---|---|
DHPMA | 120 | 6.38 |
MBA | 80 | 4.26 |
AIBN | 2 | 1 (of monomers) |
DMSO | 888 | 89.4 |
1,4-butanediol | 395 | |
Dodecanol | 397 |
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Gil, J.; Passalacqua, G.; Deloche, A.; Vidal, F.-X.; Dugas, V.; Demesmay, C. Optimization of the Preparation of Hydrophilic Poly(DHPMA-co-MBA) Monolithic Capillary Columns: A New Support for Affinity Chromatography. Separations 2023, 10, 437. https://doi.org/10.3390/separations10080437
Gil J, Passalacqua G, Deloche A, Vidal F-X, Dugas V, Demesmay C. Optimization of the Preparation of Hydrophilic Poly(DHPMA-co-MBA) Monolithic Capillary Columns: A New Support for Affinity Chromatography. Separations. 2023; 10(8):437. https://doi.org/10.3390/separations10080437
Chicago/Turabian StyleGil, Julie, Gaëtan Passalacqua, Adrien Deloche, François-Xavier Vidal, Vincent Dugas, and Claire Demesmay. 2023. "Optimization of the Preparation of Hydrophilic Poly(DHPMA-co-MBA) Monolithic Capillary Columns: A New Support for Affinity Chromatography" Separations 10, no. 8: 437. https://doi.org/10.3390/separations10080437