How do I measure nonspecific binding in an LC system for its impact on peak shape in SEC of proteins? - WKB72318

OBJECTIVE or GOAL

Measure protein interactions with the LC system materials in flow path to understand the impact on protein peak shape.

ENVIRONMENT

• Size exclusion chromatography (SEC)
• Proteins
• Nonspecific binding

PROCEDURE

1. Dilute the sample if it is concentrated, or buffer-exchange the sample into the mobile phase to be used by the method.
2. Put a union in place of the column.
3. Observe peak shape. (Depending on the system, a fast, 20/sec data collection rate or a data rate adjustment will be needed to pick up the peak as it moves through the detector.)

ADDITIONAL INFORMATION

For additional information, see Size Exclusion and Ion-Exchange Chromatography of Proteins Using the ACQUITY UPLC H-Class System.