Abstract
Background:
TOYOPEARL MX-Trp-650M is a mixed-mode resin (Tosoh Bioscience, Japan), which mediates both cation exchange and hydrophobic interactions. While mixed-mode resins are generally effective at removing antibody aggregates, reports specifically evaluating the application of MX-Trp-650M for this purpose remain limited. A previous study suggested that effective separation of monomeric and aggregated antibodies using MX-Trp-650M was achieved only under dual-gradient elution with pH and salt.
Objective:
This study aimed to further evaluate MX-Trp-650M's aggregate separation potential under various elution conditions.
Methods:
In an antibody purification case where aggregates were the predominant byproducts, both Capto MMC ImpRes and MX-Trp-650M were evaluated as the first polishing step following Protein A capture. Aggregate separation was monitored and assessed using native gel electrophoresis and size-exclusion chromatography-high-performance liquid chromatography.
Results:
MX-Trp-650M marginally outperformed Capto MMC ImpRes and achieved excellent aggregate clearance under either salt or pH mono-gradient elution.
Conclusion:
Combining the results from prior studies, the current data confirm the strong aggregate separation capability of MX-Trp-650M. The results also suggest that the optimal elution conditions for efficacious separation may vary across different antibody purification scenarios.
Keywords:
Aggregate; Antibody; Capto MMC ImpRes; MX-Trp-650M; Mixed-mode resin; Salt gradient; pH gradient.