TSKgel BioAssist S
Specially designed for the separation of large biomolecules such as antibodies, the large pores (130 nm) of the TSKgel BioAssist S cation exchange columns offer superior capacity and resolution at a low column pressure drop. They are constructed via a polymerization technique that allows an equivalent density of sulfopropyl ionic exchange groups to be incorporated into the particle without reducing pore size.
The standard TSKgel BioAssist S column is 4.6 mm ID x 5 cm, while a 10 mm ID x 10 cm semi-preparative column is available for scale-up. Both columns are made of PEEK to reduce protein adsorption.
Analysis of bromelain
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Columns: A: TSKgel BioAssist S, 7 µm, 4.6 mm ID × 5 cm
B: Competitor S, 5 mm ID × 5 cm
Mobile phase: 20 min (TSKgel) or 30 min (Competitor S) linear gradient of NaCl from 0 to
0.5 mol/L in 20 mmol/L sodium phosphate buffer, pH 7.0
Flow Rate: 0.8 mL/min for TSKgel; 1.0 mL/min for Competitor S
Detection: UV @ 280 nm
Temperature: 25 °C
Sample: crude bromelain (C4882, Sigma), 1 mg in 100 µL
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Separation of IgM by cation exchange chromatography
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Column: TSKgel BioAssist S, 7 µm, 4.6 mm ID × 5 cm
Mobile phase: 20 mmol/L sodium phosphate buffer, pH 6.0
Gradient: 0 mol/L - 0.3 mol/L NaCl (5 min) 0.3 mol/L - 0.5 mol/L NaCl (10 min)
Flow Rate: 1 mL/min
Detection: UV @ 280 nm
Sample: 500 µL of 9.5 mg/mL IgM in mouse ascites fluid; shaded peaks represent
albumin and IgM respectively |
Analysis of protein standards
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Columns: A: TSKgel BioAssist S, 7 µm, 4.6 mm ID × 5 cm
B: Conventional S type product C, 5.0 mm ID × 5 cm
C: Conventional S type product D, 4.6 mm ID × 5 cm
Mobile phase: A: 20 mmol/L sodium phosphate buffer, pH 6.5
B: 20 mmol/L sodium phosphate buffer containing 1.0 mol/L NaCl, pH 6.5
Gradient: 32 min (A-B)
Flow Rate: 0.8 mL/min
Detection: UV @ 280 nm
Temperature: 10 °C
Injection vol.: 20 µL
Samples: 1. myoglobin, 1 g/L
2. α-chymotrypsinogen A, 2 g/L
3. ribonuclease A, 4 g/L
4. cytochrome C, 2 g/L
5. lysozyme, 2 g/L |
Analysis of peptides
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Column: TSKgel BioAssist S, 7 µm, 4.6 mm ID × 5 cm
Mobile phase: A: 20 mmol/L sodium acetate buffer, pH 5.0
B: 20 mmol/L sodium acetate buffer containing 1.0 mol/L NaCl, pH 5.0
Gradient: A to B linear gradient (20 min)
Detection: UV @ 280 nm
Temperature: 25 °C |