Empowering ADC Development: Welch Materials Launches a New Key Tool for DAR Analysis
Introduction
Antibody–drug conjugates (ADCs) play an important role in driving the advance of targeted cancer therapies. As their use expands, the need for efficient, high-precision characterization methods to guarantee safety and efficacy has never been greater.
Welch Materials’ newly launched Advanchrom Bio HIC-Butyl column, with its outstanding separation performance, provides precise, reliable, and regulatory-ready chromatographic analysis of a critical ADC quality attribute — the drug-to-antibody ratio (DAR) — supporting the accelerated translation of innovative ADC candidates into the clinic and market.
Product Information
Advanchrom Bio HIC-Butyl is a high-performance column designed for protein separation based on hydrophobic interaction chromatography (HIC). It utilizes an advanced macroporous silica matrix combined with a unique surface bonding technique.
By precisely optimizing the thickness of the hydrophilic layer and the butyl bonding density on the silica surface, it is well-suited for the separation of antibodies, antibody-drug conjugates (ADCs), and other protein-based biopharmaceutical products.
Compared to Advanchrom HIC-Butyl, Bio HIC-Butyl has a weaker retention.
Product Features
- Unique chemical design offering excellent selectivity for ADC molecules
- Ultra-high purity large-pore silica microsphere matrix for superior column efficiency
- Low non-specific adsorption ensuring high recovery rates
- Excellent pressure tolerance and durability
- Outstanding batch-to-batch reproducibility
Product Specifications
| Column | Advanchrom Bio HIC-Butyl |
| Matrix | Ultra-high purity large-pore silica |
| Particle Size | 5 µm |
| Pore Size | 1000 Å |
| Column Material | Stainless Steel |
| Dimensions | 4.6×100 mm |
| pH Range | 2.0-8.0 |
| Max Temperature | 60 °C |
| Max Pressure Tolerance | 6000 psi |
Application Examples
1. DAR Analysis of ADC Sample
Chromatographic conditions:
- Column: Advanchrom Bio HIC-Butyl (4.6×100 mm, 5 µm)
- Mobile Phases:
A: 50 mM PB solution + 2.0 M ammonium sulfate, pH=7.0
B: 50 mM PB solution, pH=7.0 : isopropanol = 75:25 - Gradient Program:
Time (min) A% B% 0 90 10 1 90 10 25 0 100 30 0 100 - Flow Rate: 1.0 mL/min
- Wavelength: 280 nm
- Column Temperature: 30 °C
- Volume: 5 µL
- Sample: Disitamab Vedotin (ADC reference formulation)
2. DAR Analysis of Cysteine conjugated ADC
Chromatographic conditions:
- Column: Advanchrom Bio HIC-Butyl (4.6×100 mm, 5 µm)
- Mobile Phases:
A: 50 mM PB solution + 1.5 M ammonium sulfate, pH=7.0
B: 50 mM PB solution, pH=7.0 : isopropanol = 80:20 - Gradient Program:
Time (min) A% B% 0 100 0 20 0 100 25 0 100 - Flow Rate: 1.0 mL/min
- Wavelength: 280 nm
- Column Temperature: 30 °C
- Volume: 10 µL
- Sample: Cysteine conjugated ADC
3. Determination of Pembrolizumab
Chromatographic conditions:
- Column: Advanchrom Bio HIC-Butyl (4.6×100 mm, 5 µm)
- Mobile Phases:
A: 100 mM PB solution + 2.0 M ammonium sulfate, pH=7.0
B: 100 mM PB solution, pH=7.0 - Gradient Program:
Time (min) A% B% 0 90 10 10 90 10 15 0 100 20 0 100 - Flow Rate: 1.0 mL/min
- Wavelength: 280 nm
- Column Temperature: 30 °C
- Volume: 10 µL
4. Determination of Standard Protein Mixture
Chromatographic conditions:
- Column: Advanchrom Bio HIC-Butyl (4.6×100 mm, 5 µm)
- Mobile Phases:
A: 100 mM PB solution + 2.0 M ammonium sulfate, pH=7.0
B: 100 mM PB solution, pH=7.0 - Gradient Program:
Time (min) A% B% 0 90 10 10 90 10 15 0 100 20 0 100 - Flow Rate: 1.0 mL/min
- Wavelength: 280 nm
- Column Temperature: 30 °C
- Volume: 10 µL
- Samples: 1. Myoglobin; 2. RNase A; 3. Lysozyme; 4. α-Chymotrypsin
Welch Advanchrom Bio HIC-Butyl achieves good retention and separation among 4 standard proteins, with excellent batch-to-batch reproducibility, ensuring reliable analysis results.
Quality Assurance
Each Advanchrom Bio HIC-Butyl column is manufactured with a mature packing process and strict quality control to ensure superior separation performance and batch reproducibility.
Ordering Information
| P/N | Product | Specification |
|---|---|---|
| 0004-05039 | Advanchrom Bio HIC-Butyl | 5 µm, 4.6×100 mm |