Acoustophoresis mediated chromatography processing: Capture of proteins from cell cultures

Abstract

Chromatographic purification of target biomolecules is an important downstream process step in the development of new therapeutic agents, such as antibodies. This step employs chromatographic processes, such as affinity separation utilizing Protein A, ion exchange, or mixed mode chemistries. The workflow typically involves a packed column(s) and several chromatographic steps to achieve the desired level of purity. The steps can be time-consuming and expensive. A new process will be described, employing an acoustic standing wave in a fluid chamber to partition and maintain solid phase beads in an acoustically fluidized bed format to capture, wash, and elute the target biomolecule. Purification workflow(s) will be described for the following applications: (1) capture of a monoclonal antibody by Protein A beads from a crude cell culture system, (2) capture of recombinant Green Fluorescent protein (GFP) by anion exchange from a crude cell lysate. The workflow will include wash step(s) and recovery with a specific elution conditions. This communication will clearly demonstrate that an acoustophoresis process can purify proteins without a packed chromatography column. This new approach will minimize the time and cost involved in current purification workflows.