Mixed Mode Chromatography
New biotherapeutics, including nanobodies, bispecific mAbs, and fusion proteins, present challenges to traditional downstream purification platforms. These novel biopharmaceutical targets require specific selectivities to achieve product purity. Additionally, higher mAb titers and newly engineered mAb-like fusion proteins drive the need for increased productivity and selectivity in purification processes. Mixed mode chromatography combines multiple interaction types (e.g., ionic, hydrophobic, affinity) on a single ligand to overcome these challenges. These features:
- Enable multimodal separation to tackle tough targets
- Deliver high purity and selectivity in a single step
- Reduce process complexity and improve productivity
Mixed Mode Chromatography Portfolio
Find the Right Solution for Your Chromatography Process
Mixed mode chromatography combines multiple interaction types, including anion exchange (AEX), cation exchange (CEX), hydrophobic interaction (HIC), and affinity chromatography, on a single ligand to improve separation efficiency and reduce process steps. Explore our mixed mode solutions to find the right fit for your purification challenges.
Unique Selectivity with Mixed Mode Resins
Achieve superior separation of complex proteins with mixed mode resins.
| Resin | MEP HyperCel | HEA HyperCel | PPA HyperCel | CMM HyperCel | HA Ultrogel® |
|---|---|---|---|---|---|
| Modes | AEX & HIC | AEX & HIC | AEX & HIC | CEX & HIC | Affinity & CEX |
| Specificity | Hydrophobic | Hydrophobic | Very Hydrophobic | Hydrophobic | Selectivity for vaccines (pertussis) & recombinant proteins |
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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.
| CIMmultus® PrimaS | CIMmultus® H-Bond | |
|---|---|---|
| Channel sizes | 2 µm | 1.3, 2 µm |
| Column volume | 1 m-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 |
| Request a Quote | Request a Quote |
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
- 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
Supporting Products & Services
Learn More About Mixed Mode Chromatography
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Proof of Concept for MAbs Purification: From Batch to Intensified and Connected Process
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Benefits of Mixed-Mode Cation Exchange Chromatography: CMM HyperCel Resin Used for Monoclonal Antibody and Other Recombinant Protein Purification
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Efficient Contaminant (HCP, DNA) Removal Using MEP HyperCel Chromatography Resin for Monoclonal Antibody Purification
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Single-Domain Antibody Capture with MEP & CMM HyperCel
Sartorius mixed mode resins offer enhanced selectivity by combining ion exchange, hydrophobic interaction, and affinity chromatography. These resins support a broad range of pH and ionic strength conditions without requiring pre- or post-diafiltration steps, enabling more streamlined, intensified processes.
This study demonstrates the efficacy and versatility of MEP HyperCel as a capture platform for a wide variety of single-domain antibodies (sdAbs). sdAbs, also known as VHH fragments or nanobodies, are camelid-derived heavy-chain antibodies with unique structural and functional properties. Unlike full-length mAbs, which can be easily purified using Protein A affinity chromatography, sdAbs lack a universal binding domain, making their purification more complex and highly dependent on tailored solutions.
MEP HyperCel offered:
- Effective capture of sdAbs with neutral to acidic isoelectric points and varying molecular sizes
- High dynamic binding capacity (DBC) and recovery
- Strong impurity clearance, including host cell DNA and protein
CMM HyperCel also proves to be a valuable option, particularly for acidic sdAbs requiring near-neutral pH elution, offering flexibility in process optimization.
Further refinement of binding conditions at low salt concentrations, along with optimized contact times and DBC, may allow even greater process efficiency using the MEP and CMM HyperCel capture platform. This work was performed in collaboration with VALIDOGEN, a leading contract R&D organization (CRO) specializing in recombinant protein expression using its proprietary UNLOCK PICHIA® expression system.
Frequently Asked Questions
“Mixed mode” or “multimodal” refers to a resin’s ability to interact with molecules through multiple binding mechanisms simultaneously. The majority of Sartorius mixed mode resins combine ion exchange (IEX) and hydrophobic interaction chromatography (HIC) to enhance separation performance. The mechanism separates biomolecules first by their isoelectric point and then by their relative hydrophobicity, providing an additional dimension of selectivity.
Mixed mode resins act as a “safety belt” in downstream processing when:
- Your process lacks affinity resins (e.g., for alternative mAb platforms)
- Co-eluting species are difficult to separate or remove
- Target molecules are structurally similar, making resolution challenging
- Molecules are sensitive to buffer conditions and require narrow pH or conductivity ranges
Value of Mixed Mode Resins:
- Accelerate time to clinic by removing multiple impurities (e.g., aggregates, HCP, charge variants) in fewer steps
- Address the demands of complex upstream mixtures
- Offer unique selectivities not found in other mixed mode resins
- Ideal for applications with strict purity requirements
- The rigid cellulose matrix offers high chemical robustness, high porosity, low non-specific binding, and excellent flow properties at low backpressur
