Shodex SEC Replacements for TSKgel G2000SWXL in Protein Method Development

Record: 692

Category: Column Selection
Product: KW-802.5 and KW402.5-4F
Sample: Proteins

Question

A customer is using a TSKgel G2000SWXL size-exclusion column for protein analysis but sees broad, tailing peaks when separating a target protein from aggregates. They would like to know which Shodex SEC column corresponds to G2000SWXL and whether there is a Shodex option that can improve resolution and/or shorten the analysis time.

Answer

The closest Shodex equivalent to TSKgel G2000SWXL is PROTEIN KW-802.5. TSKgel G2000SWXL has a calibration range of 5,000–150,000 Da, and KW-802.5 covers a comparable region (5,000–100,000 Da), making it the appropriate one-to-one replacement for transferring an existing method. Shodex applications with KW-802.5 demonstrate typical SEC performance for proteins within this molecular-weight range:

Shodex applications using KW-802.5 show typical performance for standard proteins in this range:

• Standard Proteins (1) – PROTEIN KW-802.5
• Standard Proteins (14) – PROTEIN KW-802.5

If the customer requires sharper peaks, improved monomer/aggregate resolution, or shorter run times, Shodex recommends upgrading to the high-efficiency semi-micro column PROTEIN KW402.5-4F. KW402.5-4F uses smaller particles and an optimized pore structure, providing substantially higher efficiency than KW-802.5. It is especially advantageous when the target protein falls within its optimal MW range (approximately 2–70 kDa), where the column delivers excellent monomer/aggregate separation and faster elution.

Relevant applications include:

• Comparison Between KW402.5-4F and KW-802.5
• Standard Proteins (24) – PROTEIN KW402.4-4F

These studies demonstrate that KW402.5-4F provides greater resolution and reduced analysis time, outperforming conventional 8.0 × 300 mm SEC columns such as KW-802.5 or G2000SWXL.

In addition to column selection, increasing the salt concentration in the SEC mobile phase (for example, using a higher ionic-strength phosphate or NaCl buffer) can often improve peak shape and resolution by suppressing secondary interactions between the protein and the silica surface.

For final column selection, it is important to confirm the target protein’s molecular weight, the proportion of aggregates present, the level of resolution required between monomer, dimer, and higher–molecular-weight species, and the current SEC buffer and flow conditions. These details ensure that the recommended pore size and column format provide the best balance of resolution, analysis speed, and method robustness. Please contact us at support@shodexhplc.com with this information so we can recommend the most suitable SEC column for your application.

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