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Mixed-Mode Chromatography
What is mixed-mode chromatography?
Mixed-mode chromatography is a versatile separation technique that utilizes multiple modes of separation to achieve greater selectivity and specificity in separating complex mixtures of biological molecules. It combines the principles of both ion exchange and hydrophobic interactions to achieve efficient separation and purification of complex mixtures.
Unlike traditional chromatography methods that solely rely on a single interaction mechanism, mixed-mode chromatography employs a unique stationary phase that contains both hydrophobic and charged functional groups. This allows for a broader range of interactions with the target molecules, resulting in enhanced selectivity and increased resolution. By utilizing this combined approach, mixed-mode chromatography offers a powerful tool for scientists to separate and purify diverse compounds, making it an invaluable tool in the field of biotechnology for drug development and protein purification.
POROS Caprylate Mixed-Mode Cation Exchange Chromatography Resin
Thermo Scientific POROS Caprylate Mixed-Mode Cation Exchange Chromatography Resin is designed to remove moderate to high levels of aggregates during downstream processing of therapeutic antibodies in flow-through mode. As a mixed-mode, weak cation-exchange resin, POROS Caprylate effectively removes other impurities, such as host cell proteins and a wide range of high molecular weight species.
Features and benefits
Features:
- Selective capture of aggregates and hydrophobic high-molecular-weight impurities
- Flow-through design that helps save buffer and material costs
- Excellent scalability and high productivity
- Wide operational range
- Non-animal-derived
Key benefits:
- Handles high levels of aggregates
- Can reduce 10% aggregation to < 1-2% in a single polish step
- >80-90% mAb monomer recovery with minimal process optimization
- Flow-through mode increases productivity and can reduce cost of goods by 2-5X compared to bind and elute operation
Applications
A platform for purification of all biologics
Figure 1. An example of a typical downstream purification process for antibodies and proteins
Resin design
POROS and Caprylic Acid form a mixed-mode, hydrophobic weak cation exchange resin
The unique resin backbone consists of crosslinked poly(styrene-divinylbenzene) with a superior pore structure that provides rapid mass transport and facilitates increased productivity. The particle surface is coated with a proprietary hydrophilic polymer, which is further derivatized with caprylic acid.
Figure 2: 50-micrfon bead graphic
Resin performance
Results- DoE study
Click image to enlarge
Figure 3: design space for monomer vs. aggregate percentage
Results- Load density study
Click image to enlarge
Figure 4: Monomer recovery (dark blue) vs aggregate accumulation (orange), with aggregate levels marked for 1%, 2%, and 3%
Featured asset
Technical poster
Learn more about wider operational space, which allows for effective removal of aggregates and challenging molecules with high levels of aggregates, in this scientific poster.
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Bioprocess Resin Selection Tool
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Pharmaceutical Grade Reagent. For Manufacturing and Laboratory Use Only.