Columns

Hydrophobic Interaction Chromatography Column
BioPro HIC HT

BioPro HIC HT is a hydrophobic interaction chromatography column based on a non-porous hydrophilic polymer. It is suitable for analysis of biopharmaceuticals such as antibody-drug conjugates (ADC’s). Optimized surface modification and column packing condition realize high pressure tolerance and high separation ability. Therefore, high-throughput analysis with a faster flow rate can be achieved.

SPECIFICATION
Matrix Hydrophilic non-porous polymer
Particle size 2.3 µm
Bonded phase Butyl group
Usable temp. range 10-60°C
Usable pH range 2-12
Pressure limit 20 MPa (33 X 4.6 mmI.D.)
40 MPa (100 X 4.6 mmI.D.)
Features

Ideal for high resolution and rapid analysis of ADC

BioPro HIC HT consists of the fine non-porous polymer particle designed for separation of proteins, and has high pressure tolerance with excellent chromatographic resolution by optimized packing condition. This is capable of the rapid analysis without losing the resolution at a high pressure, which is too high to use with the other commercially available HIC column.

Column 100 X 4.6 mmI.D.
Eluent

A) 20 mM NaH2PO4-Na2HPO4 containing 1.0 M (NH4)2SO4 (pH 7.0)
B) 20 mM NaH2PO4-Na2HPO4 (pH 7.0)/2-propanol (85/15)

Temperature 25°C
Detection UV at 280 nm
Injection

10 µL

Sample

Brentuximab vedotin (2.5 mg/mL)

Improving throughput with a short length column at a low pressure

Analysis throughput can be improved by using a 33 mm length column. This would contribute to rapid method development and improvement of cycle time. In addition, the column allows operation at a lower pressure.

Column BioPro HIC HT 2.3 µm
Eluent A) 100 mM NaH2PO4-Na2HPO4
  containing 2.3 M (NH4)2SO4 (pH 7.0)
B) 100 mM NaH2PO4-Na2HPO4 (pH 7.0)
Temperature 25°C
Detection UV at 280 nm
Injection 15 µL
Sample 1. Ribonuclease A (0.75 mg/mL)
2. Lysozyme (0.25 mg/mL)
3. α-Chymotrypsinogen A (0.25 mg/mL)

Innovative surface chemistry for drug-to-antibody ratio (DAR) analysis

The DAR analysis by using BioPro HIC HT and the competitor columns under two mobile phase conditions varying 2-propanol content was carried out. The competitor A column having high hydrophobicity did not elute the peak #5 completely with the mobile phase containing 2-propanol lower than 15%. The competitor B column eluted all peaks with the 10% 2-propanol mobile phase condition, but it showed the poor separation between the peaks #4 and #5. On the other hand, BioPro HIC HT eluted all peaks completely with the good separation, and another peak was partially separated on the peak #3. Additionally, it is noteworthy that the same DAR values were observed at any content of 2-propanol.

Column 100 X 4.6 mmI.D.
Eluent

A) 20 mM NaH2PO4-Na2HPO4 containing 1.0 M (NH4)2SO4 (pH 7.0)
B) 20 mM NaH2PO4-Na2HPO4 (pH 7.0)/2-propanol (90/10) or (85/15) 
0-100%B (0-15 min), 100%B (15-20 min), 0%B (20-35 min)

Flow rate 0.5 mL/min
Temperature 25°C
Detection UV at 280 nm
Injection

10 µL

Sample

Brentuximab vedotin (2.5 mg/mL)

Excellent batch-to-batch reproducibility

BioPro HIC HT exhibits an excellent batch-to-batch reproducibility. This is suitable for quality control analysis of biopharmaceuticals such as MAb’s.

Column BioPro HIC HT 2.3 µm, 100 X 4.6 mmI.D.
Eluent A) 100 mM NaH2PO4-Na2HPO4 containing 2.0 M (NH4)2SO4 (pH 7.0)
B) 100 mM NaH2PO4-Na2HPO4 (pH 7.0)
0%B (0-1 min), 0-100%B (1-11 min), 100%B (11-15 min)
Flow rate 0.5 mL/min
Temperature 25°C
Detection UV at 280 nm
Injection 15 µL
Sample 1. Adalimumab (0.5 mg/mL)
2. Trastuzumab (0.5 mg/mL)
3. Bevacizumab (0.5 mg/mL)

Ordering information

Particle size
(µm)
Column size
Inner diameter X length (mm)
Product number
2.3 4.6 X 33 BHH00SQ3-H346PTH
4.6 X 100 BHH00SQ3-1046PTH

Please contact us about the product with other sizes.