September 3, 2013

Size exclusion chromatography analysis of papain-cleaved monoclonal antibody using Superdex™ 200 Increase columns

This application note describes a simplified workflow for the production of Fab (fragment, antigen binding). Fab was produced by papain cleavage of a purified monoclonal antibody (MAb) followed by purification on HiTrap™ MabSelect SuRe™ and HiTrap Protein L columns.

Intermediates and end products were analyzed with size exclusion chromatography (SEC, also called gel filtration) on two different sized Superdex 200 Increase columns: one 300 mm and one 150 mm in length. The high resolution power of Superdex 200 Increase allows baseline separation between antibody monomer, aggregates, and fragments.

Tags: Fab; fragment antigen binding; Protein-A; MabSelect; purification; size exclusion; SEC; Superdex