Sepax Proteomix HICBu Columns

Proteomix^® HICBu columns are specially designed for high resolution and high efficiency separations of proteins, oligonucleotides and peptides. Utilizing proprietary surface technologies, Proteomix^® HICBu-NP resin is made of non-porous polystyrenedivinylbenzene (PS/DVB) beads with narrow-dispersed particle size distribution. As shown in Figure below, the PS/DVB bead is modified with butyl group that provides hydrophobic interaction with analytes. Proteomix^® HICBu-NP resin is highly rigid and mechanically stable. In comparison to silica based HIC phase media, Proteomix^® HICBu-NP phases have advantages for biomolecule separations with wide pH range (2-12) and high thermal stability. The nonporous structure and narrow particle distribution offer special selectivity, high resolution separation of proteins such as mAb (monoclonal antibody), ADC (antibody drug conjugate) and related protein fragments, DNA and oligonucleotides. Proteomix^® HICBu-NP media is applicable at laboratory discovery, laboratory-scale purification and process chromatography for the production of a few mgs to kilogram of proteins.

Proteomix^® HICBu Technical Specifications

Resin Matrix	Spherical, highly cross-linked PS/DVB
Pore Size	Nonporous
Particle Size	5 µm and 10 µm
Phase Structure	Butyl
Separation Mechanism	Hydrophobic interaction
pH Stability	2-12
Operating Temperature	Up to 80 °C
Operating Pressure	Up to 6000 psi
Mobile Phase Compatibility	Compatible with aqueous solution, a mixture of water and acetonitrile, acetone, methanol, or THF

Proteomix^® HIC columns are based on PS/DVB resin and all the surface coatings are chemically bonded onto PS/DVB support, which allows exceptional high stability. The columns are compatible with most aqueous buffers, such as ammonium sulfate, sodium acetate, phosphate, Tris and a mixture of water and acetone, methanol, acetonitrile and THF. When 25 mM sodium phosphate buffer at pH 7.0 was used as the mobile phase to run the Proteomix^® HIC, 400 injections or 3 months of usage has negligible deterioration for the columns. Since the surface chemistry is built with highly controlled process, the Proteomix^® HIC columns has high lot to lot consistency, as shown in following Figure of ADC separation from 3 lots of resins.

Conditions
Columns:	Proteomix® HICBu-NP5 (5 μm, 4.6 × 35 mm)
Mobile phase:	A) 2 M ammonium sulfate in 0.025 M sodium phosphate, pH 7.0, B) 0.025 M sodium phosphate pH 7.0, C) 100% IPA
Flow rate:	0.8 mL/min
Temperature:	25 °C
Detector:	UV 214 nm
Injection volume:	10 µL
Sample:	ADC, 1.0 mg/mL in 25 mM sodium phosphate