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Estimation of methacrylate monolith binding capacity from pressure drop data
Aleš Podgornik1, Vida Smrekar, Peter Krajnc
1Centre of Excellence for Biosensors, Instrumentation and Process Control - COBIK, Velika pot 22, 5250 Solkan, Slovenia. ales.podgornik@cobik.si
Journal of Chromatography. A
|December 25, 2012
Summary
Non-invasive characterization of convective chromatographic media using pressure drop measurements allows for accurate estimation of dynamic binding capacity (DBC) for proteins and plasmid DNA. This method simplifies quality control for monolithic supports and grafted layers.
Area of Science:
- Chromatography
- Bioseparations
- Materials Science
Background:
- Convective chromatographic media like membranes and monoliths offer fast separations for large biomolecules.
- Standard characterization methods are unsuitable for single-piece materials.
- Non-invasive techniques are preferred for characterizing these supports.
Purpose of the Study:
- To investigate pressure drop for estimating dynamic binding capacity (DBC) in monoliths with varying pore sizes.
- To explore the relationship between pressure drop and DBC in grafted layers.
- To establish a non-invasive method for quality control of convective chromatographic media.
Main Methods:
- Investigated pressure drop across methacrylate monoliths with different pore sizes.
- Analyzed DBC for proteins and plasmid DNA using ion-exchange and hydrophobic interactions.
- Applied pressure drop analysis to estimate DBC in grafted layers of varying thicknesses.
Main Results:
- Monolith surface area is inversely proportional to pore diameter.
- Pressure drop is inversely proportional to the square of pore size, indicating preserved microstructure.
- Derived a mathematical model showing DBC linearly correlates with the square root of pressure drop.
- Experimentally confirmed the DBC-pressure drop relationship for proteins and plasmid DNA.
- Demonstrated that capacity is proportional to grafted layer thickness.
Conclusions:
- Pressure drop is a reliable, non-invasive method for estimating DBC in convective chromatographic media.
- The derived mathematical formalism accurately predicts DBC based on pressure drop.
- This approach facilitates efficient quality control and process development for chromatographic supports.
