Page 82 - Shimadu Consumables Catalog

Bio LC Columns

Shim-pack Bio HIC: Hydrophobic Interaction Chromatography Column

Antibody-drug conjugate (ADC) is a next-generation antibody drug that is expected to provide strong effects in a wide range of
therapeutic treatments as compared to existing chemotherapeutic agents. The drug-to-antibody ratio (DAR) indicates the ratio of
addition of the drug that directly influences both efficacy and potential toxicity of ADCs. Analysis of DAR is extremely important
when judging important quality attributes of ADCs as it may significantly affect properties such as stability and agglutination.
Hydrophobic interaction chromatography (HIC) is suitable for separation of analytes with slightly different hydrophobic properties
such as antibody-drug conjugates (ADCs) with a different drug-to-antibody ratio (DAR). Using the Shim-pack Bio HIC, the DAR of
ADCs can be analysed with a high separation capacity.

Drug to Antibody Ratio (DAR) analysis of ADCs
mV
30 DAR4
Conditions
Column : Shim-pack Bio HIC, 4 μm, 4.6×100mm
Mobile Phase : A) 50 mM NaH PO -Na HPO (pH 7.0) containing 1.5 M
2
4
2
4
20 (NH ) SO /2-propanol (95/5)
4
4 2
B) 50 mM NaH PO -Na HPO (pH 7.0)/2-propanol (80/20)
2
2
4
4
0%B (0-1 min), 0-100%B (1-15 min), 100%B (15-20 min)
Flow Rate : 1.0 mL/min
DAR2 DAR6 Col. Temp. : 25 ºC
10 Detection : UV at 280 nm
Injection Vol. : 5 µL
DAR8
Sample : Cystein-conjugated ADC mimic (5 mg/mL)
DAR0
0
0 5 10 15 20 min
Shim-pack Bio HIC Butyl
P/N Dimensions
227-31174-01 4 μm, 100 x 4.6 mm I.D

Pg A-78