Recombinant monoclonal antibodies (MAbs) are a growing class of therapeutic molecules. Early in the development of MAbs, a large number of harvest cell culture (HCC) samples must be screened for immunoglobulin G (IgG) titer, aggregations, and charge variants. This study describes a two-dimensional (2D) high performance liquid chromatography (HPLC) workflow for afinity, size-exclusion, and ion-exchange analysis.