Solutions

Intermediate Purification

Intermediate Purification Process Step

The partially purified feedstock from the capture step will have a higher target concentration and relatively less complex impurity profile (also at higher concentrations). The intermediate purification step uses chromatography's resolving power to separate the target away from other impurities of the feedstock. Because of this, and the improved fluid flow properties, resin particle sizes of 65 µm and smaller are used. In many cases, more than one intermediate purification column may be necessary to achieve the desired degree of product purity.

The role of resin efficiency (particle size) and selectivity (ligand and pore size) become more important for intermediate purification. The question for the process chromatographer is "What percent of dynamic binding capacity can I load the resin to and still have adequate resolution from most impurities?"

Intermediate purification steps typically use smaller particles than capture steps. The viscosity of the intermediate feedstock and the operational pressure limits of the chromatographic hardware used, determine the most appropriate particle size for this process step.

Many commercial TOYOPEARL and TSKgel PW bulk resin products are offered in particle sizes from 100 µm to 20 µm and have the same methacrylic polymer backbone. The selectivity of feedstock components on these resins remains the same regardless of bead size. TOYOPEARLresins with bead diameters of 35 µm and larger are made to withstand operating pressures up to 0.3 MPa. TSKgel PW resins with bead diameters of 20 and 30 µm are made to withstand operating pressures up to 2 MPa.

 

Tosoh Bioscience has commercial products with the following particle sizes for a typical intermediate purification step:

Grade

Particle Size Range

Medium (M) 40 - 90 µm
Superfine (S) 20 - 50 µm
(30) 20 - 40 µm

 

Approximate dynamic binding capacities for typical intermediate purification resins:

Mode

Typical Protein

Antibody

Ion Exchange up to 200 mg/mL up to 150 mg/mL
Hydrophobic Interaction up to 60 mg/mL up to 45 mg/mL

Please click on the mode of interest for additional product information.

 

Chromatography Mode

Particle Size

TOYOPEARL Resin

Anion Exchange  65 µm SuperQ-650M
DEAE-650M
Anion Exchange  35 µm SuperQ-650S
DEAE-650S
GigaCap Q-650S
Cation Exchange  65 µm SP-650M
CM-650M
Cation Exchange  35 µm SP-650S
CM-650S
GigaCap S-650S
Hydrophobic Interaction  65 µm Butyl-600M
Phenyl-600M
PPG-600M

Butyl-650M
Phenyl-650M
Ether-650M
Hydrophobic Interaction  35 µm Butyl-650S
Phenyl-650S
Ether-650S
Affinity  65 µm Activated Resins
AF-Epoxy-650
AF-Tresyl-650

Reactive Resins
AF-Carboxy-650
AF-Amino-650
AF-Formyl-650

Resins with Group Specific Ligands
AF-Chelate-650
AF-Red-650
AF-Heparin HC-650