Purification Techniques | Application of MMC Series Cation Multimodal Resins in Antibody Purification

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In the downstream purification of antibody drugs (such as monoclonal antibodies, bispecific antibodies, and Fc fusion proteins), the polishing step—especially the removal of aggregates, host cell proteins (HCPs), DNA, and viruses—is crucial for product quality. Conventional cation exchange chromatography (CEX) is widely used but often faces challenges such as narrow operational windows and restricted impurity removal efficiency when handling complex samples or pursuing greater process robustness.

To address these challenges, multimodal chromatography has emerged as a vital supplement, and even an alternative, to conventional cation exchange chromatography. By combining multiple interaction mechanisms (such as electrostatic forces, hydrophobic interactions, and hydrogen bonding), multimodal chromatography allows for more efficient impurity removal under broader operating conditions while maintaining high product recovery.

This article highlights the performance characteristics and application cases of BioLink MMC series cation multimodal resins in antibody purification

Cation Multimodal Resin Series

MaXtar® MMC (average particle size: 90 μm) and MaXtar® MMC HR (average particle size: 40 μm) resins incorporate functional groups with different properties: carboxyl, hydrogen bonding, and hydrophobic moieties. Compared to traditional single-mode chromatography resins, these multimodal ligands exhibit a combination of multiple interaction forces, enabling the selective removal of diverse impurities in a single step. This results in higher recovery and superior product quality, offering significant performance advantages:

High binding capacity saves time and reduces cost; HR's smaller particles offer higher resolution.
Broad operating window for pH and conductivity in production-scale processes.
Robust MaXtar® matrix withstands high pressures, making scale-up straightforward.
With excellent salt tolerance, MaXtar® MMC is ideal for capture steps in recombinant protein processes, reducing the need for buffer exchange.
High impurity clearance efficiency, especially for aggregates, in intermediate or polishing steps of antibody purification.

Antibody Binding Capacity

Column volume: 4.7 mL
Sample: Purified IgG1 (expressed in CHO cells)
Residence time: Sample loading duration: 5 min
Binding buffer: 50 mM HAc-NaAc, pH 5.5

purification-techniques-application-of-mmc-series-cation-multimodal-resins-in-antibody-purification1.jpg

Figure 1: MaXtar® MMC Antibody Binding Capacity

Summary:Under ionic-binding mode, average binding capacity: MaXtar® MMC: 50 mg/mL; MaXtar® MMC HR: 60 mg/mL

Salt Tolerance Characterization

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Figure 2: MaXtar® MMC Salt Tolerance Test

Summary: Under ionic-binding mode, when using 50 mM HAc–NaAc, pH 5.5 buffer and progressively increasing NaCl concentrations, IgG remained well bound up to 150 mM NaCl(17 mS/cm).

Pressure-Flow Curve

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Figure 3: MaXtar® MMC Pressure-Flow Curve

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Figure 4: MaXtar® MMC HR Pressure-Flow Curve

Summary: Both MaXtar® MMC and MaXtar® MMC HR show excellent mechanical performance. The finer-particle HR variant maintains performance suitable for commercial-scale applications while delivering higher resolution.

Application Case 1: MaXtar® MMC HR Efficient Removal of Aggregates and Fragments in an Antibody Project

Test Conditions:

Resin: MaXtar® MMC HR
Sample: monoclonal antibody sample (15 mg/mL)
Buffer A: 50 mM NaAc-HAc, pH 5.5
Buffer B: 20 mM PB, 0.5 M NaCl, pH 7.0

Sample	SEC purity			NE-CE purity
Sample	HMW (%)	Monomer (%)	LMW (%)	HMW (%)	Monomer (%)	LMW (%)
Load	6.43	79.58	13.99	5.55	75.58	18.87
Elution	0.45	98.43	1.12	0.00	98.11	1.89

Table 1: MaXtar® MMC HR Process Data

Summary: After purification with MaXtar® MMC HR, aggregate content dropped from 6.43 % to 0.45 %, and fragments decreased from 13.99 % to 1.12 %. The results demonstrate that MaXtar® MMC HR provides a clear and effective removal of both aggregates and fragments.

Application Case 2: MaXtar® MMC HR Removal of Acidic/Basic Peak in an Antibody Purification

Test Conditions:

· Resin: MaXtar® MMC HR, 4.7 mL column

· Load: Sample at 3 mS/cm, pH 5.5 (<20 mg/mL)

· Buffer A: 50 mM NaAc–HAc, pH 5.5

· Buffer B: 50 mM NaAc–HAc + 1 M NaCl, pH 5.5

purification-techniques-application-of-mmc-series-cation-multimodal-resins-in-antibody-purification5.jpg

Figure 5: Elution Profile of MaXtar® MMC HR

purification-techniques-application-of-mmc-series-cation-multimodal-resins-in-antibody-purification6.jpg Figure 6: Acidic/basic Peak Detection Chromatogram of MaXtar® MMC HR

Sample	Yield (%)	Charge Variants
Sample	Yield (%)	Acidic Peak (%)	Main Peak (%)	Basic Peak (%)
Load	/	45	53	2
Elution	78	33	66	1

Table 2: MaXtar® MMC HR Process Data

Summary: The results show that MaXtar® MMC HR effectively removes acidic charge variants. Acidic peak elute first, followed by basic peak.

Selection Guide & Practical Tips

MaXtar® MMC has an average particle size of 90 μm, while MaXtar® MMC HR is 40 μm. In addition to particle size differences, MMC HR features a lower ligand density than MMC, resulting in weaker binding strength. If overly strong binding is observed during process development, MMC HR is recommended.
For highly hydrophobic samples, if elution cannot be achieved effectively using standard salt or pH conditions with the MMC series, a combined salt/pH gradient is recommended.
When the sample contains high levels of product-related impurities such as aggregates, fragments, or charge variants, the higher resolution of MaXtar® MMC HR is recommended.
Process development should be guided by the specific properties of the sample and purification goals. Special attention should be paid to achieving a balance between product purity and recovery yield.

References

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