Development of BioRad NGC and GE ÄKTA pure systems for highly automated three column protein purification employing tandem affinity, buffer exchange and size exclusion chromatography

Publication date: Available online 7 September 2019Source: Protein Expression and PurificationAuthor(s): Dwight Winters, Mai Tran, Daniel Yoo, Kenneth WalkerAbstractAffinity purification, such as Protein A (Pro A) followed by size exclusion chromatography (SEC) remains a popular method to obtain research scale proteins. With the need for higher throughput protein production increasing for discovery research, there is substantial interest in the automation of complex protein purification processes, which often start with a Pro A step followed by SEC. However, the harsh elution conditions from Pro A based chromatography can destabilize some proteins resulting in particulates, which in turn can cause column fouling and potential cross-contamination of subsequent purifications. We modified both Bio Rad NGC and ÄKTA Pure systems to run a three-column process (Pro A to buffer exchange to SEC) enabling automated tandem affinity to SEC purification while minimizing the risk of SEC column fouling and subsequent cross-contamination. The intervening buffer exchange column, unlike the final SEC column, can be rapidly regenerated using harsh methods between runs, and these automated systems are capable of processing up to six samples per day without user intervention.

Source: Protein Expression and Purification - September 8, 2019 Category: Biochemistry Source Type: research