Biological therapeutics continues to be a highly researched area within the pharmaceutical and biotechnology industry.  With the growing success of monoclonal antibody based drugs on the market, significant work is now focused on enhancing the targeted approach of these compounds through the development of Antibody Drug Conjugates (ADC’s) or bispecific antibodies.<br/>ADC’s are generally created by modifying a native mAb in way which allows for linkage to a cytotoxic payload.  Bispecific antibodies or fusion proteins tend to be formed by bringing together two protein fragments or domains from two different proteins to form a non-native antibody species.  In the case of bispecific antibodies, this is typically performed through a technique referred to as Fab-arm exchange, in which the Fab-arm, or half-IgG fragment of one antibody is attached to the Fab-arm of another antibody.   One of the challenges of such an approach is removing intact antibodies from the formed Fab-arm in order to obtain a high yield Fab-arm exchange.<br/>Here, we show the effectiveness of separating the Fab-arm from the intact antibody and low molecular weight fragment with high resolution using Size Exclusion Chromatography (SEC) and other orthogonal chromatographic modes.  Additionally, the use of SEC in the characterization of the PEGylated Fab-arm from the native species is also evaluated.  The results shown here are expected to be highly applicable in the development of bispecific antibodies as protein therapeutics.

Characterization of native and PEGylated Fab-arms using various modes of analytical chromatography

249th National Meeting (2015)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.