This Application Note describes results in protein impurity testing and simultaneous molecular mass characterization with the Agilent 1100 Series GPC-SEC analysis system.
This document details the use of aqueous size exclusion chromatography (SEC), also known as gel filtration chromatography, for analyzing proteins. It highlights the Agilent 1100 Series GPC-SEC analysis system as a tool for impurity testing, reaction monitoring, and molecular mass determination without denaturing the protein.
The note presents experimental results demonstrating the system's ability to separate and quantify protein impurities, such as dimers, alongside accurate molecular weight characterization. It compares the SEC method with other techniques like SDS-PAGE and mass spectrometry, emphasizing the benefits of automation and the ability to collect samples for further investigation.
The technique can handle a very wide range of molecular weights, from several hundred to about 10 million Dalton.
Molecular weight accuracy and precision are typically better than 2 %.
A polydispersity D value smaller than 1.01 typically indicates peak purity.