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.

Full Scalability and Support

Matrices scalable from benchtop to commercial manufacturing


 

Unique Selectivities

Unique mixed-mode ligands offer increased selectivity and resolution, offering a more powerful separation for complex purification

High Productivity

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.


Resins

Monoliths

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

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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.

CMM HyperCel

MEP HyperCel

HEA HyperCel

PPA HyperCel

HA Ultrogel®

Modes

CEX

HIC

Hydrophobic charge
induction (HCIC)
pseudo-affinity
to IgG (thiophilic)

AEX

HIC

AEX

HIC

CEX

Pseudo-affinity

Application

B/E antibody
HCP, aggregate removal
recombinant molecule purification

Capture of antibodies in physiological conditions (pseudo affinity) only for MEP HyperCel
HCP, aggregate, variants, removal
antibody, fab, enzymes, recombinant proteins

 

Dynamic binding capacity

IgG
>60-100 mg/mL*

Human IgG
>20 mg/mL*

BSA
40-60 mg/mL*

BSA
40-60 mg/mL*

Cytochrome C
>7 mg/mL**

Average particle size (µm)

50-80

80-100

80-100

80-100

60-180

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* 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.

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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.

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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.

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PPA HyperCel

PPA HyperCel exhibits a stronger hydrophobicity with an aromatic ligand, allowing protein binding at a moderate salt concentration under neutral pH.

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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.

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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.


CIMmultus® PrimaS

CIMmultus® H-Bond

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

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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

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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

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Consult Our Experts

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Biomolecule Analysis Tools

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