TSKgel UP-SW Aggregate
TSKgel UP-SW Aggregate, the latest addition to the UP-SW series of silica-based size exclusion chromatography (SEC) columns, are specifically designed to facilitate the analysis of mAb aggregates, high molecular weight (MW) proteins and nucleic acids. These columns are 3 µm, 30 nm pore size SEC analytical columns that have been designed with a higher exclusion limit than other TSKgel UP-SW columns.
Available in 4.6 mm ID, TSKgel UP-SW Aggregate columns are compatible with both UHPLC and HPLC systems and require less sample while delivering higher sensitivity compared to the 7.8 mm ID TSKgel UltraSW Aggregate column. These columns feature high pore volume per unit column volume, low sample adsorption (due to the derivatization of the particle surface with ligands containing diol functional groups) and excellent column efficiency, all contributing to unsurpassed sample resolution.
Analysis of mAb aggregates
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Columns: A: TSKgel UP-SW Aggregate, 3 µm, 4.6 mm ID × 30 cm
B: TSKgel UP-SW3000, 2 µm, 4.6 mm ID × 15 cm
Mobile phase: 40 mmol/L sodium phosphate buffer, pH 6.7 + 400 mmol/L sodium perchlorate
+ 0.05% sodium azide
Flow rate: 0.20 mL/min
Detection: UV @ 280 nm
Temperature: 25 °C
Injection vol.: 10 µL
Sample: mAb HMWs: dimer, aggregates; LMWs: fragments |
Analysis of heat forced mAb aggregation
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Column: TSKgel UP-SW Aggregate, 3 µm, 4.6 mm ID × 30 cm
Mobile phase: 40 mmol/L sodium phosphate buffer, pH 6.7, + 400 mmol/L sodium perchlorate
+ 0.05 % sodium azide
Flow rate: 0.2 mL/min
Detection: UV @ 280 nm
Temperature: 25 °C
Injection vol.: 10 μL Sample: mAb
* The sample was diluted by 10-fold with 20 mmol/L sodium phosphate buffer (pH 7.2) + 150 mmol/L sodium chloride and dispensed to small aliquots. Each aliquot was stored at following temperature respectively for 2 hours. 70 °C (blue chromatogram)
73 °C (red chromatogram)
77 °C (green chromatogram)
80 °C (light blue chromatogram) |