HPLC - Affinity

1. What is the support material for Tosoh Bioscience affinity resins?

TSKgel affinity chromatography columns are based upon the G5000PW methacrylate porous resin. This “5PW” media is hydrophilic, with 100 nanometer (nm) pores and an estimated protein size exclusion limit of 1000 kilodaltons.

2. What are the ligand concentrations, adsorption capacities and test analytes for the various affinity column supplied by Tosoh Bioscience?

Characteristics of TSKgel AFC Columns   
Column Packing  Ligand Type  Ligand Concentration  Adsorption Capacity  Sample 
Boronate-5PW  m-aminophenyl boronate not available  40 µmol/mL - resin  sorbitol
Chelate-5PW  iminodiacetic acid  20 µmol/mL - resin  not available not available
Tresyl-5PW  tresyl   ca. 20 µmol/mL - resin > 60 mg/g dry resin (coupling capacity)   soybean trypsin inhibitor


3. Can I get a TSKgel Affinity LC column with non-metal wetted parts?

Yes. We supply glass columns in two formats: 5 mm ID x 5 cm L and 8 mm ID x 7.5 cm L with 10 µm pore size Pyrex frits.

The shipping solvents for these columns are as follows:

Boronate-5PW: distilled water
Chelate-5PW: acetate buffer
Tresyl-5PW: acetone


4. How do I load and desorb protein from a TSKgel affinity column?

Specific instructions for retaining your protein as well as desorption procedures are included with each affinity column. Keep in the mind that the analyte requires some structural feature that coincides with the retention mechanism of the affinity resin.

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