TSKgel® HIC-ADC Butyl – 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-Butyl column sets 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
Due to their technical performance, these columns simplify method development and tech-transfer processes for DAR analyses. In addition, the TSKgel HIC-ADC-Butyl columns enhance both the accuracy and efficiency of Quality Control processes and enable the highest quality standards in ADC product.
Specifications and technical details
| Base material: |
Polymeric (polymethacrylate) |
| Pore size (mean): |
Non-porous |
| Separation mechanism: |
Hydrophobic interaction chromatography (HIC) |
| Functional group: |
Butyl |
| 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 column
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 column was benchmarked against other commercially available HIC columns. The TSKgel HIC-ADC Butyl column showed superior separation of DARs and their isoforms.
Separation performance of TSKgel HIC-ADC Butyl column in comparison to other commercial HIC columns
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An ADC mimic was analyzed by HIC-ADC using the following conditions:
Columns: all 4.6 mm ID x 10 cm L
TSKgel HIC-ADC Butyl (P/N 0023539)
Non-Tosoh HIC columns as indicated
Mobile phases: Buffer A: 1.5 mol/L (NH₄)₂SO₄+ 0.1 mol/L PBS pH 7.0 + 5% IPA
Buffer B: 0.1 mol/L PBS pH 7.0 + 20% IPA
Gradient: 0 min (0%B) 1 min(0%B) → 30min (100%B)
Flow rate: 0.5 mL/min
Detection: UV @ 215 nm
Inj. vol.: 10 μL
Sample: 0.5 mg/mL SigmaMAb Antibody Drug Conjugate (ADC) Mimic (Merck MSQC8)
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