On-line purification of monoclonal antibodies using an integrated stirred-tank reactor/expanded-bed adsorption system.

While expanded-bed adsorption (EBA) units have been used to recover proteins from whole cell cultures, the development of a more efficient, on-line process could streamline the traditional multistep process. This study implements a bench-scale on-line purification system in which whole cell cultures are loaded directly into a chromatography column to capture a monoclonal antibody (mAb) in a single step. The on-line purification system used here integrates a stirred-tank reactor (STR) and an EBA unit into a new hybrid (STR-EBA) system.

Determination of monoclonal antibody production in cell culture using novel microfluidic and traditional assays.

This study compares microfluidic technology (Protein 200 LabChip Assay kit, Agilent 2100 Bioanalyzer, referred to here as Protein 200) to the traditional approach for protein analysis, one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), for the sizing and quantification of immunoglobulin G (IgG) in hybridoma cell cultures. Internal references differ between each method: purified IgG was used alone in SDS-PAGE while myosin (the upper marker) was added to each sample in Protein 200. The IgG used here were produced in cultures propagated in either a serum-free or a serum-containing medium.