TSKgel G3000SWXL size exclusion HPLC column for proteins and mAbs
The G3000SWXL is a size exclusion chromatography (SEC) column compatible with all HPLC systems. With a pore size of 25 nm, the column is suited to analyze proteins between 10-500 kDa and has become the industry standard for the analysis of aggregates and fragments in protein biotherapeutics and especially monoclonal antibodies.The column belongs to the group of SWXL columns (silica-based SEC columns for aqueous conditions), it is also available for larger (TSKgel G4000SWXL) and smaller proteins (TSKgel G2000SWXL).
Specifications and technical details
| Base material |
Silica |
| Particle size (mean) |
5 µm |
| Pore size (mean) |
25 nm |
| Separation mechanism |
gel filtration chromatography (GFC)/
aqueous size exclusion chromatography |
| Calibration range |
1.0 × 104 - 5.0 × 105 Da (globular proteins) |
| Functional group |
Diol |
| pH stability |
2.5-7.5 |
| Salt stability |
≤ 0.5 mol/L |
| Organic concentration |
0 – 100% |
| Temperature |
0-30 °C |
| Max. Flow rate |
1.2 mL/min (standard: 0.5-1.0 mL/min) |
| USP column compatibility |
USP L59, USP L20 |
Calibration curves for TSKgel G3000SWXL compared to other SWXL columns
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Separation of a protein standard on TSKgel SWXL columns (7.7 mm ID x 30 cm L).
Mobile phase: 0.3 mol/L NaCl in 0.1 mol/L sodium phosphate buffer pH 7.0
Detection: UV @ 220 nm
Sample: 1. Thyroglobulin (660 kDa) 2. IgG (160 kDa) 3. BSA (67 kDa) 4. Ovalbumin (43 kDa)
5. Peroxidase (40 kDa) 6. Beta-lactoglobulin (18 kDa) 7. Myoglobulin (17 kDa)
8. Ribonuclease A (13 kDa) 9. Cytochrome C (12 kDa) 10. Glycine tetramer (0.25 kDa)
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Benefits of the G3000SWXL – reproducibility, resolution, range of products
The SWXL size exclusion columns are valued for their high reproducibility. Furthermore, they exhibit a high lifetime and good resolution due to a high pore volume as well as low sample adsorption. In addition to the classical column with a stainless steel housing of 7.8 mm ID and 30 cm length, the column is further available in 15 cm length for fast analyses (TSKgel QC-PAK GFC 300) or packed in a PEEK housing for highest biocompatibility (TSKgel BioAssist G3SWXL).
Application of the G3000SWXL column - aggregate and fragment analysis
The TSKgel G3000SWXL column is used in quality control (QC) and during development processes to analyze the content of aggregate and fragment impurities, also referred to as high molecular weight impurities (HMW) and low molecular weight impurities (LMW). Even though it is used for various protein biotherapeutics, the main analyte is monoclonal antibodies (mAbs). Upon injection onto the column, the larger aggregates will elute first while the monomer elutes slightly later, followed by the smaller fragments. This way, the percentage of high- and low-molecular weight impurities as well as the monomer purity can be determined which needs to be within product-specific limits to pass QC. An example of an antibody separation is given in the application note “TSKgel G3000SWXL Columns for the reproducible analysis of mAbs and proteins”
Monoclonal antibody analyzed on a G3000SWXL size exclusion column
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Column: TSKgel G3000SWXL, 5 µm, 7.8 mm ID × 30 cm
Instrument: Agilent 1260
Mobile phase: 0.2 mol/L potassium phosphate/0.25 mol/L KCl, pH 6.2
Flow rate: A. 0.5 mL/min
Detection: UV @ 280 nm
Injection: 50 µL |