Mixed-Mode Chromatography
New biotherapeutics such as nanobodies, bispecific mAbs and fusion proteins, present new challenges to traditional downstream purification platforms. Additionally, higher mAb titers and newly engineered mAb-like fusion proteins now drive the need for higher productivity/selectivity purification.
Sartorius Mixed-Mode (MM) chromatography ligands interact with target protein molecules through multiple types of interactions such as affinity, hydrophobic interaction (HIC), and cation/anion exchange. These multimodal separation mechanisms allow resins to be applied to solve challenging purification problems where traditional IEX, HIC, or affinity chromatography alone do not meet the need.
Matrices scalable from benchtop to commercial manufacturing
Unique mixed-mode ligands offer increased selectivity and resolution, offering a more powerful separation for complex purification
Plug-and-play chromatography matrices allow high-speed processing without sacrificing capacity
Find the Right Answer for Your Chromatography Process
Novel biopharmaceutical targets such as nanobodies, multi-specific mAbs , and fusion proteins require new selectivities to achieve high product purity. Mixed-mode matrices leverage multimodal separation mechanisms such as IEX and HIC to achieve higher selectivity and purity in a single step compared to traditional single-mode chromatography.
Application | Capture and polish small and medium biomolecules | Capture and polish large biomolecules |
Implementation | Multi-use | Intrabatch multi-use, single-use between batches |
Ease-of-use | Bulk material (easy packing columns) | Ready-to-use |
Scalability | Wide range of column sizes | Wide range of device sizes |
Matrices | CMM, HEA, PPA, MEP, HA | H-bond ADC, PrimaS |
Mixed-Mode Resins
New Purification Challenges Require a Different Approach
Novel biopharmaceutical targets such as nanobodies, plural-specific mAbs, and fusion proteins require unique selectivities to achieve high product purity. Mixed-mode resins leverage multi-modal separation mechanisms such as IEX and HIC to achieve higher selectivity and purity in a single step than in traditional single-mode chromatography.
Sartorius offers a broad range of selectivities to address challenges with a high concentration of contaminants with the intensified feedstreams, aggregates issues with newer modalities, and to remove challenging process or product-related impurities. Achieve reduced process costs by streamlining pre- or post-diafiltration steps.
Modes | CEX HIC | Hydrophobic charge | AEX HIC | AEX HIC | CEX Pseudo-affinity |
Application | B/E antibody | Capture of antibodies in physiological conditions (pseudo affinity) only for MEP HyperCel; HCP, aggregate, variants, removal |
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Dynamic binding capacity | IgG | Human IgG | BSA | BSA | Cytochrome C |
Average particle size (µm) | 50-80 | 80-100 | 80-100 | 80-100 | 60-180 |
* 10% BT
** 50% BT
CMM HyperCel
CMM HyperCel is a hydrophobic cation exchanger recommended for the capture of antibodies, antibody fragments, and recombinant proteins. It allows the separation of proteins with similar pI by modulation of pH and conductivity.
MEP HyperCel
MEP HyperCel provides unique selectivity for monoclonal antibody capture or purification to remove aggregates, HCP, and DNA. The ligand facilitates the binding of target proteins at neutral pH under moderate salt concentrations.
HEA HyperCel
HEA HyperCel is a mixed-mode ligand with a less hydrophobic hexylamine chain allowing protein binding at a moderate salt concentration under neutral pH.
PPA HyperCel
PPA HyperCel exhibits a stronger hydrophobicity with an aromatic ligand, allowing protein binding at a moderate salt concentration under neutral pH.
HA Ultrogel®
HA Ultrogel® hydroxyapatite is a composite material of cross-linked agarose and microcrystalline hydroxyapatite enclosed in the agarose matrix. The material shows mixed-mode functionality based on cation exchange and metal affinity in the hydroxyapatite structure. It is ideally suitable for general impurity removal.
Mixed Mode Monolithic Columns
Unique Selectivity with Mixed Mode Monolithic Columns
The unique selectivity of mixed-mode monolithic columns ensures the proper purification of the most challenging large biomolecules.
Channel sizes | 2 µm | 1.3, 2 µm |
Column volume | 1 mL – 8 L | 1 mL – 8 L |
Working range, pH | 2 – 13 | 2 – 13 |
Functionality | Hydrogen bonding and anion exchange | Multimodal hydrogen donor-acceptor |
Application | mRNA | Viruses and extra-cellular vesicles |
CIMmultus® PrimaS
Multimodal chromatography ligand that combines elements of hydrogen bonding with anion exchange chromatography
Unique selectivities in many important product areas
Supports purification of single-stranded RNA at ambient temperature
Fractionates ssRNA by size
CIMmultus® H-Bond ADC
Multimodal hydrogen donor-acceptor
Working range, pH 2–13
Enables size-based separation of product and contaminants at acidic pH
Elution with increasing salt, pH, or sorbitol
Highly tolerant of salt
Outstanding DNA removal
Unique selectivity for preparative and analytical applications
Sanitizable with 1 M NaOH
