SOLUTIONS

TSKgel UP-SW3000-LS

UHPLC size exclusion column for protein analysis by light scattering and mass spectrometry

The TSKgel UP-SW3000-LS columns are dedicated to UHPLC size-exclusion analyses of monoclonal antibodies and other biopharmaceuticals combined with advanced detection such as multi-angle light scattering (MALS) or mass spectrometry (MS). The 2 µm silica particles are diol bonded for a hydrophilic surface. In combination with the 25 nm pore size, the columns are optimally suited for the analysis of proteins ranging between 10 kDa – 500 kDa. The column is also available without improvements for advanced detection methods: TSKgel UP-SW3000.

Protein Calibration Curves for UP-SW3000-LS SEC column

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Column:           TSKgel UP-SW3000-LS, 2 µm, 4.6 mm ID × 30 cm
                         TSKgel UP-SW3000, 2 µm, 4.6 mm ID × 30 cm
                        Competitor SEC, 1.7 µm, 4.6 mm ID × 30 cm
Mobile phase:  100 mmol/L phosphate buffer, pH 6.7 + 100 mmol/L Na2SO4
Flow rate:         0.35 mL/min
Detection:        UV @ 280 nm
Temperature:   25 °C
Inj. vol.:            10 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

Noise of buffer injection in multi-angle light scattering detection

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fig4-web-up-sw-Ls.png

Column:           TSKgel UP-SW3000-LS, 2 µm, 4.6 mm ID × 30 cm
                        Competitor SEC, 1.7 µm, 4.6 mm ID × 30 cm
Mobile phase:  100 mmol/L phosphate buffer, pH 6.7 + 100 mmol/L Na2SO4
Flow rate:         0.35 mL/min
Detection:        Low angle light scattering (LALS) detected with LenS3 MALS detector
Temperature:   25 °C
Inj. vol.:            10 μL
Samples:          Mobile phase only

Key feature of UP-SW3000-LS UHPLC columns is a highly reduced noise level in advanced detectors saving time for equilibration and improving data quality. For multi-angle light scattering (MALS) applications, a low noise level is achieved in minutes, leading to high signal-to-noise ratios and thus high sensitivity. For mass spectrometry applications, ionization efficiency by electrospray ionization is improved and increases overall MS performance and instrument uptime. This makes the UP-SW3000-LS column particularly suited for in-depth analyses of aggregates and fragments of mAbs, bi- and multispecifics.

TSKgel UP-SW3000-LS lot-to-lot reproducibility

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Column:           TSKgel UP-SW3000-LS, 2 µm, 4.6 mm ID × 30 cm of three different production lots
Mobile phase:   100 mmol/L phosphate buffer, pH 6.7 + 100 mmol/L Na2SO4
Flow rate:         0.35 mL/min
Detection:        UV @ 280 nm
Temperature:   25 °C
Inj. vol.:            10 μL
Samples:           Protein standard containing thyroglobulin, gamma-globulin, ovalbumin, ribonuclease A, p-amino
                          benzoic acid, 137 Da

In addition, the UP-SW3000-LS columns offer reproducible and reliable size exclusion protein separations. This is explained by robust separations over multiple injections as well as a high column lifetime which reduces the need for frequent column replacements. However, low variations between column production lots facilitate column exchange.

 

Base Material: Silica
Particle Size (mean): 2 µm
Pore Size (mean): 25 nm
Functional Group: Diol
pH Stability: 2.5 - 7.5
Calibration Range (globular proteins): 10,000 - 500,000 Da

Ordering Information ​

P/N Description Dimension
0023546 TSKgel UP-SW3000-LS 4.6 mm ID x 15 cm L
0023547 TSKgel UP-SW3000-LS 4.6 mm ID x 30 cm L
0023548 TSKgel guardcolumn UP-SW-LS 4.6 mm ID x 2.0 cm L
0023549 TSKgel guardcolumn UP-SW-LS DC 4.6 mm ID x 2.0 cm L