Mixed Mode Chromatography

Mixed-Mode Chromatography

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. It is a functionalized version of the TOYOPEARL® HW size exclusion resin and is therefore based on a hydroxylated polymethacrylic polymer bead. TOYOPEARL® MX-Trp-650M resin uses tryptophan as the active ligand. This amino acid 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.

Structure of TOYOPEARL® MX-Trp-650M


In addition to fast binding and elution kinetics (see below), 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.


Maintains DBC post-CIP with 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)

DBC Measurement
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 (1.0 mL/min)
Detection: UV @ 280 nm
Sample: polyclonal human IgG
Sample load: 1 mg/mL


Excellent Elution Kinetics


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
Sample load: 1 mg/mL diluted with buffer A



Pore size (mean) 100 nm Visit the TOYOPEARL® MX-Trp-650M product page 
Particle size (mean) 75 µm 
Pressure rating 0.3 MPa
Shipping buffer  20% ethanol
pH stability 3-13
Shelf life 10 years (estimated)