Mixed-Mode Chromatography | Thermo Fisher Scientific

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

Bioprocessing workflow image showing a typical downstream purification process for antibodies and proteins.

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.

Graphic illustrating hydrophobic weak cation exchange resin

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.

Download poster