Increasing Throughput of a Tandem mAb Purification by Adding a High-Volume Autosampler to the NGC™ Chromatography System
Monoclonal antibodies (mAbs) have become an important and highly effective class of biopharmaceuticals.To identify mAbs with ideal specificity, affinity, and binding kinetics, hundreds of candidates have to be purified to high purity and screened in downstream functional assays. We describe a highly robust and reproducible, fully automated mAb purification process that allows purification of 84 samples in two days, yielding milligram quantities of highly pure protein. Using this method we have purified approximately 500 mAb drug candidates by eliminating the protein purification throughput bottleneck.
Monoclonal antibodies (mAbs) are used in the clinic to treat a variety of diseases, including various cancers, multiple sclerosis, and rheumatoid arthritis. To develop a single therapeutic mAb hundreds of candidates are purified and screened. The downstream functional assays used to characterize candidate mAbs require milligram quantities of highly pure protein in appropriate buffers. Low pH, for example, can lead to mAb degradation, and cell-based functional assays require specific salt concentrations and pH. These stringent requirements have made mAb purification a bottleneck in the discovery process.
To increase the throughput of our existing mAb purification protocol we coupled an NGC Discover™ Pro Chromatography System to a Teledyne CETAC ASX-560 Autosampler. The NGC™ Discover Pro System's multidimensional (Multi-D) chromatography function allows automated multicolumn purification workflows while the ASX-560 Autosampler accommodates up to eighty-four 50 ml conical tubes for automatic sample loading. The NGC Signal Import Module (SIM) was used to facilitate communication between the two instruments.
We show that, using this medium-throughput automated process, 84 samples can be purified in two days, yielding pure protein in a highly reproducible manner.