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Empowering ADC Development: Welch Materials Launches a New Key Tool for DAR Analysis

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.
Structure of antibody-drug conjugates

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.

Advanchrom Bio HIC-Butyl

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
An Advanchrom Bio HIC-Butyl column

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)
DAR distribution of disitamab vedotin sample
DAR distribution of disitamab vedotin sample

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
DAR distribution of cys-ADC
DAR distribution of cys-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
Chromatogram of pembrolizumab (Keytruda)
Chromatogram of pembrolizumab (Keytruda)

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
Chromatograms of standard proteins from 3 batches
Chromatograms of standard proteins from 3 batches

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