SOLUTIONS

TSKgel UP-SW3000

TSKgel UP-SW3000 

TSKgel UP-SW3000 silica-based GFC columns are packed 2 μm particles, providing high efficiency UHPLC separations of proteins. The 25 nm pore size results in a fractionation range up to 500,000 Da for globular proteins. These UHP-SEC columns are ideally suited for the analysis of antibodies, their fragments, and aggregates in biopharmaceutical R&D or QC analysis. They feature the same pore size as TSKgel SWXL, the current industry’s gold standard for QC analysis of therapeutic antibodies. Thus, the new UP-SW UHPLC columns allow a smooth method transfer from HPLC to UHPLC.

It is important to employ an UHPLC system to take full advantage of the high efficiency that can be obtained on TSKgel UP-SW columns. The columns can also be used with an HPLC system that is optimized with regards to extra-column void volume but maximum plate numbers will not be achieved with HPLC instruments.

Protein calibration curve for TSKgel UP-SW3000 and competitor SEC column 

upsw_fig1.png 

Column:          TSKgel UP-SW3000, 2 µm, 4.6 mm ID × 15 cm
                         Competitor SEC, 1.7 µm, 4.6 mm ID × 15 cm
Mobile phase: 100 mmol/L phosphate buffer, pH 6.7
                         + 100 mmol/L Na2SO4 + 0.05% NaN3
Flow rate:        0.35 mL/min
Detection:        UV @ 280 nm
Temperature:  25 °C
Inj. vol.:           5 μL
Samples:          1. thyroglobulin, 640 kDa
                          1’. thyroglobulin dimer
                          2. gamma-globulin, 155 kDa
                          2’. gamma-globulin dimer
                          3. ovalbumin, 47 kDa
                          4. ribonuclease A, 13,700 Da
                          5. p-amino benzoic acid, 137 Da
 

 

Comparison of Antibody Analysis Results 

upsw_fig2.png 

mouse-human chimeric mAb
1: trimer; 2: dimer; 3: monomer ; 4: fragment

Column  RS (peak 1/2)  RS (peak 2/3) 
TSKgel UP-SW3000 2 μm 1.52  3.56 
Competitor UHPLC-SEC 1.7 μm  1.25  3.47 

 

TSKgel UP-SW3000 column-to-column reproducibility 

upsw_fig3.png 
Column:          TSKgel UP-SW3000, 2 µm, 4.6 mm ID × 30 cm
Instrument:    Thermo Fisher/Dionex Ultimate 3000 UHPLC System
Mobile phase: 100 mmol/L sodium phosphate buffer, pH 6.7, +
                         100mmol/L Na2SO4 + 0.05% NaN3
Gradient:        Isocratic
Flow rate:       0.35 mL/min
Detection:       UV @ 280 nm
Temperature: 25 °C
Injection vol.: 5 µL
Samples:         QC standard protein test mixture:
                         thyroglobulin, 640 kDa, 0.5 g/L
                         gamma-globulin, 155 kDa, 1 g/L
                         ovalbumin, 47 kDa, 1 g/L
                         ribonuclease A , 13,700 Da, 1.5 g/L
                         p-aminobenzoic acid, 137 Da, 0.01 g/L 
Base material  Silica
Particle size (mean) 2 μm 
Pore size (mean)  25 nm
Functional group  Diol
pH stability  2.5 - 7.5
Calibration range (proteins)  10-500 kDa 

 

Ordering Information

P/N Description Dimension
0023449 TSKgel UP-SW3000 4.6 mm ID x 15 cm L
0023448 TSKgel UP-SW3000 4.6 mm ID x 30 cm L
0023450 TSKgel Guardcolumn UP-SW3000 4.6 mm ID x 2.0 cm L
0023451 TSKgel DC Guardcolumn UP-SW3000 4.6 mm ID x 2.0 cm L