Mixed-Mode or Multimodal Chromatography expands the range of chromatographic modes applied in biopurification. Mixed-Mode media combine ionic and hydrophobic interactions and offer new selectivities and a higher salt tolerance than traditional ion exchange media.
Mixed-Mode Resins are especially suited for the purification of target molecules that are difficult to purify using common purification platforms. Mixed-mode media can be used for direct processing of clarified feedstocks at physiological salt concentrations as well as for intermediate and polishing applications, such as mAb aggregate removal.
TOYOPEARL Mixed-Mode Resins
TOYOPEARL MX-Trp-650M is a high capacity mixed-mode resin. This resin uses tryptophan as the active ligand and is based on a hydroxylated polymethacrylic polymer bead. The amino acid tryptophan has both weak carboxyl cation exchange and indole hydrophobic functional groups. The selectivity of the resin can be adjusted through control of binding or elution pH, ionic strength, salt type and additives. This multi-modal resin maintains dynamic binding capacity (DBC) at elevated feedstock and buffer conductivities.
In addition to fast binding and elution kinetics (see figure), TOYOPEARL MX-Trp-650M minimizes the amount of WFI needed for either feedstock dilution during capture or column elution further downstream, leading to lower in-process pool volumes and higher throughput.
TOYOPEARL MX-Trp-650M maintains a high DBC even after extended CIP (clean in place) cycles using 0.5 mol/L NaOH, as demonstrated in the figure below.
Structure of TOYOPEARL MX-Trp-650M
Stability in 0.5 mol/L NaOH
|Resin: TOYOPEARL MX-Trp-650M |
Alkaline cleaning (CIP) conditions
3CV: 0.5 mol/L NaOH,
5CV: 0.1 mol/L Tris-HCl, pH 8.5 + 0.3 mol/L NaCl
Flow rate: 212 cm/hr (1.0 mL/min)
Column size: 6 mm ID × 4 cm
Binding buffer: 0.05 mol/L acetate buffer, pH 4.7 + 0.1 mol/L NaCl
Flow rate: 212 cm/hr (1 mL/min)
Detection: UV @ 280 nm
Sample: polyclonal human IgG
Sample Load: 1 mg/mL
Narrow elution peak widths
|Resin:TOYOPEARL MX-Trp-650M |
Column size:6 mm ID × 4 cm
Mobile phase:Buffer A: 0.05 mol/L acetate buffer, pH 4.7 + 0.1 mol/L NaCl (12 mS/cm)
Buffer B: 0.1 mol/L Tris-HCl buffer, pH 8.5 + 0.3 mol/L NaCl
Flow rate:A: 212 cm/hr (1.0 mL/min)
B: 424 cm/hr (2.0 mL/min) started at 124 min
Detection:UV @ 280 nm
Sample:CHO cell culture media, monoclonal antibody (1 mg/mL) diluted with buffer A