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Technical Support
(877) SEPAX-US, press 3  TechSupport@sepax-tech.com |
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Showcases
Customer Showcase 1:
SEC Column Phase Screening Service
Sample:
Monoclonal antibody F, MW 145 kD, pI=8.4
Goal:
In search of high resolution separation between protein monomer, aggregates, and fragments.
Approach:
Screen different column phases (Zenix-300 and Zenix-C 300) to achieve best separation.
Conclusion:
Zenix-C SEC-300 (3 µm, 300 Å) column delivers baseline separation of monoclonal antibody monomer and aggregates.
Customer Showcase 2:
IEX Column Phase Screening Service
Sample:
Monoclonal antibody F, MW 145 kD, pI=8.4
Goal:
In search of most suitable stationary phase and separation conditions for the analysis of antibody F variants.
Approach:
Screen variety of cation exchange stationary phases (Proteomix SCX, WCX and Antibodix WCX) at analytical level (4.6 × 250 mm) with mobile phase screening.
Conclusion:
Antibodix NP5 (5 µm, 4.6 × 250 mm) column with NaCl salt gradient at pH 7.5 offers the best selectivity and resolution for the antibody variant separation among the phases screened.
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LiCl salt gradient Column: Proteomix SCX-NP5, WCX-NP5 and ABX WCX-NP5 Mobile phase: A: 20 mM NaAc, adjust pH to 5.15 by 380 µL HAc, B: A + 1 M LiCl Flow rate: 0.8 mL/min Detector: UV 280 nm Column temperature: 30 °C Injection volume: 10 µL Pressure: 79 bar |  |
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NaCl salt gradient pH 7.5 Column: Proteomix SCX-NP5, WCX-NP5 and ABX WCX-NP5 Mobile phase: A: 20 mM Phosphate buffer, pH 7.5; B: A + 1 M NaCl Flow rate: 0.8 mL/min Detector: UV 280 nm Column temperature: 30 °C Injection volume: 10 µL Pressure: 79 bar |  |
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pH/salt gradient Column: Proteomix SCX-NP5, WCX-NP5 and ABX WCX-NP5 Mobile phase: A: 2.4 mM Tris, 1.5 mM Imidazole, 11.6 mM piperazine, adjust pH to 6.0 by 10 M NaOH; B: A + 0.5 M NaCl, adjust pH to 10.5 by 10 M NaOH Flow rate: 0.8 mL/min Detector: UV 280 nm Column temperature: 30 °C Injection volume: 10 µL Pressure: 79 bar |  |
Customer Showcase 3:
SEC Mobile Phase Screening Service
Sample:
CHO cell expressed Fc Fusion Protein, 160 kD
Goal:
In search of high resolution separation between protein monomer, dimer, and high molecular weight aggregates.
Approach:
Screen different mobile phase to achieve best separation and sample recovery.
Conclusion:
Zenix SEC-300 (3 µm, 300 Å) column delivers baseline separation of Fc fusion protein, its oligomer, and impurity. With the addition of arginine, the hydrophobic interaction between Fc fusion protein and column resin is minimized.