NEW 1.7 µm  Proteomix® UHPLC HIC Column 
Hydrophobic Interaction Chromatography (HIC) is an efficient method to monitor and isolate protein variants including species due to isomerization, oxidation, aggregation, degradation. For example, in mAb stability studies, oxidation of exposed amino acid residues such as tryptophan and methionine may affect its safety, efficacy, and may impact product shelf life and bioactivity. 

Sepax Proteomix® HIC 1.7 µm offers high resolution separation of mAb oxidized variants. The small 1.7 µm particle size allows for a highly sensitive way to capture and monitor the small changes of variants. It can be used as an  orthogonal method to SEC and IEX, when they are not sufficient to characterize low level variants with minimum size or charge difference. 

Because of the mild non-denaturing running conditions, native protein molecule structures are preserved when using HIC. The  heterogeneous mAb variants can be fractionated by  Proteomix® HIC  prep columns into different groups based on their hydrophobicities, which then can be subjected to other techniques such as IEX, RP, peptide mapping or potency assay for further characterization. 
1

2 Hour Oxidation

1.7 µm vs 5  µm
Column: Proteomix® HIC Butyl NP1.7 µm, (1.7 µm, 4.6 x 100 mm), PN: 431NP2-4610 Mobile phase: A: 100 mM sodium phosphate buffer, 2 M ammonium sulfate, pH 7.0,B: 100 mM sodium phosphate buffer, pH 7.0 Flow rate: 0.8 mL/min, Detector: UV 214 nm, Column temperature: 25 C; Pressure: 560 bar Injection: 12.5 µg mAb sample, 25 µg oxidized mAb; Check more details on the application note
Product Information:

Proteomix® HIC Butyl NP 1.7 µm
4.6 X 35 mm (P/N: 431NP2-4603)
4.6 X 100 mm (P/N: 431NP2-4610)

Proteomix® HIC Butyl NP 5 µm
4.6 X 35 mm (P/N: 431NP5-4603)
4.6 X 100 mm (P/N: 431NP5-4610)

*2.1, 7.8, 10, 21.2, 30 mm I.D. sizes are also available. Check for more details here.
 

Downloads

Full Application Note of 
mAb Oxidation on HIC 1.7 µm 


MAb Hydrophobicity Screening


Column: Proteomix® HIC Butyl NP 1.7 (1.7 µm, 4.6 x 35 mm);  Mobile phase: A: 100 mM sodium phosphate buffer, 2 M ammonium sulfate, pH 7.0, B: 100 mM sodium phosphate buffer, pH 7.0;  Flow rate: 0.8 mL/min, Detector: UV 214 nm, Column temperature: Ambient; Injection:  10 µg mAb samples, 1 mg/mL Erbitux, 0.5 mg/mL Herceptin in 1 M ammonium sulfate, 50 mM phosphate buffer, 2.5 mg/mL rituximab in 500 mM ammonium sulfate, 25 mM phosphate buffer
Column: Proteomix® HIC Butyl NP5 (5 µm, 4.6 x 35 mm);  Mobile phase: A: 100 mM sodium phosphate buffer, 2 M ammonium sulfate, pH 7.0, B: 100 mM sodium phosphate buffer, pH 7.0, C: B + 1 M NaCl;  Flow rate: 1 mL/min, Detector: UV 214 nm, Column temperature: Ambient; Injection: 15 µg mAb samples; Pressure: 62-70 bar
CONTACT US
www.sepax-tech.com · 1-877-SEPAX-US