Development of an automated mid-scale parallel protein purification system for antibody purification and affinity chromatography

Publication date: December 2016 Source:Protein Expression and Purification, Volume 128 Author(s): Chi Zhang, Alexander M. Long, Brooke Swalm, Ken Charest, Yan Wang, Jiali Hu, Craig Schulz, Wolfgang Goetzinger, Brian E. Hall Protein purification is often a bottleneck during protein generation for large molecule drug discovery. Therapeutic antibody campaigns typically require the purification of hundreds of monoclonal antibodies (mAbs) during the hybridoma process and lead optimization. With the increase in high-throughput cloning, faster DNA sequencing, and the use of parallel protein expression systems, a need for high-throughput purification approaches has evolved, particularly in the midsize range between 20 ml and 100 ml. To address this we modified a four channel Gilson solid phase extraction system (referred to as MG-SPE) with switching valves and sample holding loops to be able to perform standard affinity purification using commercially available columns and micro-titer format deep well blocks. By running 4 samples in parallel, the MG-SPE has the capacity to purify up to 24 samples of greater than 50 ml each using a single-step affinity purification protocol or a two-step protocol consisting of affinity chromatography followed by desalting/buffer exchange overnight (∼12 h run time). Our evaluation of affinity purification using mAbs and Fc-fusion proteins from mammalian cell supernatants demonstrates that the MG-SPE compared favorably with industry sta...

Source: Protein Expression and Purification - August 10, 2016 Category: Biochemistry Source Type: research