TSKgel® HIC-ADC columns – for antibody-drug-conjugate (ADC) analysis
The name gives it away: TSKgel HIC-ADC columns are hydrophobic interaction chromatography columns designed for the analysis of Antibody-drug conjugates (ADCs). Tosoh has developed a new particle construction to achieve exceptional separation of variably conjugated ADC variants. This next-generation breakthrough in non-porous particle technology makes DAR analysis more reliable and improves separation efficiency. The TSKgel HIC-ADC columns set a new industry standard with respect to the following characteristics.
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- Superior separation performance
- Fast analyses (e.g. DAR determination in 7 minutes)
- Reliability and lifetime
- Ligand selection for diverse molecules: Phenyl (high hydrophobicity),
Butyl (mid hydrophobicity)
The columns simplify method development and tech-transfer processes for DAR analyses, enhancing both the accuracy and efficiency of Quality Control processes, and enabling the highest quality standards in ADC analysis.
Specifications and technical details
| Base material: | Polymeric (polymethacrylate) |
| Pore size (mean): | Non-porous |
| Separation mechanism: | Hydrophobic interaction chromatography (HIC) |
| Functional group: | Butyl, Phenyl |
| pH stability: | 2.0-12.0 |
| Organic concentration: | 0 – 50% |
| Temperature: | 10-60 °C |
| Dimensions: | 4.6 mm ID × 3.5 cm L and 4.6 mm ID × 10 cm L,
guard column available (4.6 mm ID x 5 mm L) |
Separation performance of the HIC-ADC columns
The most important prerequisite to determining the Drug-Antibody-Ratio (DAR) in ADCs is maximum resolution between different payload variants. Individual peak areas can then be determined and used to calculate the average DAR. The columns were benchmarked against other commercially available HIC columns, as well. The TSKgel HIC-ADC Butyl column demonstrates the superior resolution particularly of high DARs while the TSKgel HIC-ADC-Phenyl column is particularly suited for low hydrophobic species.
Separation performance of TSKgel HIC-ADC columns in comparison to other commercial HIC columns. |
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An ADC mimic was analyzed by HIC-ADC using the following conditions: | Columns | TSKgel HIC-ADC Phenyl, TSKgel HIC-ADC Butyl, Commercial HIC Columns (4.6 mm I.D. × 10 cm) | | Mobile Phase A | 0.1mol/L Phosphate Buffer pH 7.0 +1.0 mol/L Ammonium Sulfate | | Mobile Phase B | 0.1 mol/L Phosphate Buffer pH 7.0 / Isopropanol (70/30, v/v) | | Gradient | 0 min (0%B), 1 min (0%B) → 15 min (100%B) | | Flow rate | 0.5 mL/min | | Detection | UV @ 215 nm | | Temperature | 25℃ | | Injection volume | 20 μL | | Sample | 0.5 mg/mL ADC Mimic (Sigma) | |