This article will demonstrate a new protocol for streamlining the traditional recombinant protein purification workflow by combining the enzymatic lysis and purification steps. This approach results in significantly less hands-on time and greater than two-hour time savings over the traditional workflow ( Figure 1). Incorporation of magnetic beads into the protocol eliminates the need to clarify lysates by centrifugation. Magnetic beads have been adopted where agarose beads have been used, reducing processing time and increasing sample throughput. Magnetic beads are generally used in batch mode and isolated on a magnet to allow for buffer exchange. Magnetic beads facilitate automation of the protocol using a particle processor, resulting in increased sample throughput while reducing hands-on processing time to less than 10 minutes. To further improve the workflow, we used BugBuster® Master Mix reagent (EMD Millipore), which allows for nonmechanical extraction of soluble protein from bacterial cells.
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