Mixed-Mode Chromatography Resins

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Support purification of complex targets with mixed-mode chromatography resins

Achieving purity targets in biologic purification workflows can be challenging. Mixed-mode chromatography (MMC), also referred to as multimodal chromatography, combines multiple interactions, such as ionic and hydrophobic interactions, within a single functional group to improve selectivity in complex feedstreams. This multiple-interaction approach can enhance selectivity and offers a robust, scalable solution for meeting product quality targets.

Benefits of POROS Caprylate Mixed-Mode Cation Exchange Resin

The Thermo Scientific POROS Caprylate Mixed-Mode Cation Exchange Chromatography Resin is designed for high-efficiency flow-through polishing in antibody and antibody-derivative workflows for process intensification. Benefits include:

  • Suitable for high aggregate levels in monoclonal antibody (mAb), bispecific, and multispecific antibody processes
  • Capable of reducing aggregates from greater than 10% to less than 2% in a single polish step while maintaining greater than 80–90% mAb monomer recovery
  • Effective in reducing host cell proteins (HCPs), leached protein A ligand, and other product- and process-related impurities
  • Supportive of higher productivity and lower cost of goods (COGs) through efficient operation in flow-through mode
Aggregate removal with POROS Caprylate

Figure 1: Experimental example of aggregate removal with POROS Caprylate

Designed for process efficiency and high recovery

The POROS Caprylate Mixed-Mode Cation Exchange Chromatography Resin features a rigid polymer backbone and a large throughpore structure that enables rapid mass transfer and low backpressure, even at high linear velocities. The POROS base bead is functionalized with caprylic acid, which gives the resin both hydrophobic and weak cation exchange characteristics. It is designed to operate in flow-through mode over a wide range of buffer conditions. Together, these design elements help maintain high throughput, improve downstream efficiency, and support scalable purification from process development through production.

50 micron bead

Figure 2: 50-micron bead graphic

Performance data supporting scalable purification

Performance studies confirm that the POROS Caprylate Mixed-Mode Resin maintains high monomer recovery and effective aggregate reduction across a wide design space. Load density studies show greater than 80–90% monomer recovery with aggregates reduced to less than 2%, even at high loading. Design of Experiments (DoE) modeling demonstrates robust performance across varying pH and conductivity ranges.

Results—DoE study

Design space for monomer vs. aggregate percentage

Figure 3: Design space for monomer vs. aggregate percentage

Results—Load density study

Monomer recovery vs aggregate accumulation

Figure 4: Monomer recovery (dark blue) vs aggregate accumulation (orange), with aggregate levels marked for 1%, 2%, and 3%

Other modalities and applications supported

ADCs

Antibody-Drug Conjugates (ADCs) workflows rely on chromatography steps to separate complex conjugates and reduce impurities. These approaches support consistent production of ADCs for therapeutic research and biomanufacturing.
 

Cell Therapy

Cell therapies utilize living cells as treatments, necessitating strict control over inputs and processing steps. Chromatography is applied to prepare ancillary materials, such as cytokines or viral vectors, that are used during cell manufacturing.
 

Gene Therapy

Gene therapy often relies on viral vectors, such as adeno-associated virus (AAV) or lentivirus, to facilitate the transfer of genetic material. Chromatography separates drug product from product- and process-related impurities in both capture and polishing steps.

mAbs

Monoclonal antibody purification typically starts with Protein A chromatography to capture the antibody from cell culture fluid. Additional steps, such as ion exchange, are used to help reduce aggregates, variants, or process-related impurities.
 

mRNA

Chromatography in mRNA production separates full-length transcripts from double-stranded RNA and truncated products. The purified RNA can then be used in formulation or downstream steps, such as lipid nanoparticle assembly.
 

Vaccines

Chromatography is often used in vaccine manufacturing for both the capture and polishing of the vaccine drug product. The choice of method depends on the platform, for example, protein subunits, viral vectors, or nucleic acid vaccines.
 

Explore other chromatography resins

Affinity chromatography resins


Affinity resins enable capture with high purity and yield in one step. While widely used for antibody purification, CaptureSelect™ technology also supports bispecific antibodies, fragments, Fc-fusion proteins, recombinant proteins, and viral vectors using mild elution conditions.
 

Anion exchange chromatography resins

Anion exchange (AEX) resins separate biomolecules based on charge, capturing negatively charged species through interactions with positively charged groups. Strong AEX resins maintain capacity across broad pH and conductivity ranges, supporting both capture and polishing.
 

Cation exchange chromatography resins

Cation exchange chromatography (CEX) resins separate positively charged biomolecules using negatively charged functional groups on the resin matrix. POROS CEX resins offer high capacity, chemical stability, and consistent performance from development through commercial manufacturing.

Hydrophobic interaction chromatography resins

Hydrophobic interaction chromatography (HIC) separates biomolecules based on surface hydrophobicity. HIC is typically applied as a polishing step, when differences in hydrophobicity can be leveraged to improve the purification process.

 

Ion exchange chromatography resins

Ion exchange (IEX) chromatography separates biomolecules through charge-based interactions with functional groups. IEX resins are available in strong and weak base or acid chemistries to support bind/elute or flow-through purification of antibodies, vaccines, viral vectors, and other biologics.

Protein A chromatography resin


Protein A (ProA) chromatography is used for antibody purification. MabCaptureC resin offers high binding capacity, alkaline stability, and scalability while keeping costs of goods in mind, making it suitable for process development through cGMP manufacturing.
 

Explore more chromatography resins

Chromatography resins include affinity, IEX, HIC, and mixed-mode formats. They enable reproducible performance from screening tools to cGMP production, supporting process consistency across purification steps.

Not sure which resins are right for you?


Selecting the right resin depends on your molecule type, process stage, and purification goals. Compare chemistries, binding capacities, and scalability data in one place.
 

Reduce preparation time with chromatography columns

Pre-packed chromatography columns simplify method setup and reduce preparation time. Available in a range of sizes, these columns support capture, intermediate, and polishing steps across bioprocessing workflows.
 

Frequently asked questions

Mixed-mode chromatography uses ligands with multiple interaction mechanisms, such as ionic and hydrophobic, to offer superior selectivity and process control. This approach allows for fine-tuning of selectivity by helping to enhance or reduce these binding mechanisms by altering process conditions. POROS Caprylate Mixed-Mode Cation Exchange Resin, for example, uses caprylic acid’s dual interactions for the efficient reduction of aggregates and process impurities while preserving product integrity and recovery.

Mixed-mode chromatography integrates multiple interaction modes, such as ionic and hydrophobic interactions, into a single functional group. This design enables a distinct selectivity profile compared to single-mode resins, allowing for the effective separation of aggregates, variants, and impurities that may co-elute under standard ion exchange or hydrophobic interaction conditions. The combined interaction facilitates process robustness and efficient, scalable purification across a broader range of pH and conductivity settings.

The POROS Caprylate resin is designed for flow-through polishing applications in antibody and antibody-derivative purification workflows. This mixed-mode cation exchange resin is an ideal choice for feedstreams containing high levels of aggregates or where orthogonal methods for HCP reduction are desirable. The resin is designed to perform effectively at high loading densities and across a broad pH and conductivity range, supporting product quality and downstream intensification throughout development and manufacturing.

The multimodal resin operates effectively across a broad pH range typical of standard polishing conditions. In a DoE study using a Herceptin feedstock with a high percentage of high molecular weight impurities, effective and consistent aggregate reduction was found between pH 4.5 and 6. This range could change depending on the target molecule’s isoelectric point (pI) and hydrophobicity characteristics. This flexibility allows operation at higher conductivities than traditional cation exchange resins.

Mixed-mode chromatography resin samples are available. You can request a resin sample through the Biotherapeutics Sample Request Form.

Chromatography resources


Access resources that cover chromatography performance, purification strategies, and method development. These resources empower scientists to select and apply chromatography resins from research to cGMP manufacturing.

Enhance your mixed-mode chromatography process

For research use or further manufacturing. Not for diagnostic use or direct administration into humans or animals.