SOLUTIONS

TSKgel UP-SW Columns

TSKgel UP-SW Columns

The TSKgel UP-SW product line consists of three columns differing in pore size, which allows for the analysis of small to very large molecular weight biopharma products. Available in 4.6 mm ID x 15 and 30 cm lengths, TSKgel UP-SW columns are compatible with both UHPLC and HPLC systems and require less sample while delivering higher sensitivity compared to 7.8 mm ID TSKgel SEC columns.

The TSKgel UP-SW columns are:

TSKgel UP-SW2000, 12.5 nm pore size, for analysis of small proteins, peptides and oligonucleotides »

TSKgel UP-SW3000, 25 nm pore size, analysis of mAb monomer and antibody fragments »

TSKgel UP-SW Aggregate, 30 nm pore size, for analysis of very large proteins and high order mAb aggregates »

As shown in the figure, the calibration curves show differing slopes that create distinct separation ranges for each of the columns within the TSKgel UP-SW product line. The calibration curve for the TSKgel UP-SW2000 column is similar to that of the TSKgel G2000SWXL column curve, implying a better resolution of small proteins, peptides and oligonucleotides. The TSKgel UP-SW3000 column shows a shallower slope around the molecular weight region of γ-globulin, making this column ideal for the analysis of monoclonal antibodies.  The UP-SW Aggregate column demonstrates the shallowest slope of the calibration curve of proteins up to the high molecular weight region.

Calibration curves of standard proteins

calibration-curves-proteins.jpg
Column:           A. TSKgel UP-SW2000, 2 µm, 4.6 mm ID × 30 cm
                         B. TSKgel UP-SW3000, 2 µm, 4.6 mm ID × 30 cm
                         C. TSKgel UP-SW Aggregate, 3 µm, 4.6 mm ID × 30 cm
Mobile phase: 100 mmol/L sodium phosphate buffer, pH 6.7, + 100 mmol/L sodium sulfate + 0.05 % sodium azide Flow rate:        0.35 mL/min
Detection:       UV @ 280 nm
Temperature:  25 °C
Injection vol.: 10 μL
Samples:         1. thyroglobulin (MW 640,000) (1'. thyroglobulin dimer)
                         2. γ - globulin (MW 155,000) (2'. γ - globulin dimer)
                         3. ovalbumin (MW 47,000)
                         4. ribonuclease A (MW 13,700)
                         5. p - amino benzoic acid (MW 137)