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Column	BIOshell™ A400 Protein C4, 10 cm X 2.1 mm I.D., 3.4 µm particles (66825-U)
Column temp.	90 °C
Mobile phase	(A) 80:20, 0.1% TFA in water: 0.1% TFA in acetonitrile; (B) 20:80, 0.1% TFA in water: 0.1% TFA in acetonitrile
Gradient	20 to 35%B in 10 min (or 5 min)
Flow rate	0.2 mL/min
Pressure	470 psi (or 940 psi)
Sample	1 g/L mAb in 20 mM Tricine/KOH, pH 7
Injection	0.4 µL
Detector	UV, 215 nm

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Analysis Note	Antibodies can exhibit considerable hydrophobicity, and so efficient reversed-phase elution of this intact mAb requires the use of high temperature. The top chromatogram illustrates that BIOshell^™ Fused-Core^® columns can be operated at high flow rate while maintaining good peak shape at a reasonable back pressure.
Categories	Analytical Chromatography, Proteins
Featured Industry	Life Science and Biopharma
Legal Information	BIOshell is a trademark of Sigma-Aldrich Co. LLC, Fused-Core is a registered trademark of Advanced Materials Technology, Inc.
Suitability	Application for HPLC

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