Advanchrom Bio HIC-Butyl, 5µm, 4.6×100mm (1pcs/pk)

Empower ADC Characterization with High-Performance HIC Columns

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 DAR analysis method has never been greater.

Advanchrom Bio HIC-Butyl

Advanchrom Bio HIC-Butyl is a high-performance column designed for protein separation based on hydrophobic interaction chromatography. It is well-suited for the separation of antibodies, antibody-drug conjugates (ADCs), and other protein-based biopharmaceutical products.

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.

Advanchrom HIC-Butyl

Advanchrom HIC-Butyl is an ideal choice for low hydrophobicity antibodies, proteins, and ADCs.

• Achieves good retention and separation of protein samples with low ammonium sulfate concentration in the mobile phase system.
• Compatible with MS-compatible mobile phase systems (ammonium acetate, acetonitrile, etc.), correlating LC-UV separation results with LC-MS characterization more accurately and more rapidly.

Application Examples:

DAR Analysis of Disitamab Vedotin

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 Analysis of an ADC Sample

Chromatographic conditions:

• Column: Advanchrom HIC-Butyl (4.6×50 mm, 3 µm)
• Mobile Phases:
  A: 1.0 M ammonium acetate, pH=7.0
  B: 25 mM ammonium acetate : acetonitrile = 50:50
• Gradient Program:
  

  Time (min)	A%	B%
  0	100	0
  2	100	0
  20	0	100
  25	0	100

• Flow Rate: 1.0 mL/min
• Wavelength: 280 nm
• Column Temperature: 30 °C
• Volume: 10 µL

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

Determination of Standard Protein Mixture

Chromatographic conditions: Same as above

In this application, Welch Advanchrom Bio HIC-Butyl achieves good retention and separation among 4 standard proteins, with excellent batch-to-batch reproducibility, ensuring reliable analysis results.

Brand: Advanchrom

Phase: Bio HIC-Butyl

Particle Size: 5µm

Pore Size: 1000Å

USP Classification:

Separation Mode: HIC

pH Range: 2.0-8.0

Surface Area:

Carbon Load:

Max.Pressure: 6000psi

Endcapped:

Max.Temp.: 60°C

Particle Shape: Spherical

Column Cross Reference:
(Please kindly note that columns among brands are never 100% the same. The below cross reference list indicates alternatives column(s) that will likely give a similar selectivity. Inevitable differences in retention time, resolution and theoretical plate count exist and may affect the results for certain circumstances.)