Electrical asymmetrical flow field-flow fractionation analysis of the NIST monoclonal antibody reference material 8671

The electrical asymmetrical flow field-flow fractionation system allows both size and surface charge separation, enabling measurement of size or molecular weight distribution and electrophoretic mobility. The open channel design of FFF allows for better recovery of injected mass than SEC. This is important for quantifying protein/antibody aggregates present in small relative amounts, and represents an advantage of FFF over SEC for aggregate quantification.

In this work, NIST mAb is used to compare separation, aggregation, quantification, and recovery parameters for asymmetrical flow field-flow fractionation (AF4) versus size exclusion chromatography (SEC), and to demonstrate the capabilities of the electrical asymmetrical flow field-flow fractionation (EAF4) module to simultaneously measure size and surface charge characteristics of antibodies and proteins.