Transient Transfection

HTP Transient Transfection & Expressed Protein Purification

PhyNexus PhyTip Columns make it easy to implement automated workflow from transfecting mammalian cells to recovery of large volumes of high quality proteins expressed from transient transfection.  PhyTip Columns are unique in their compatibility with HTP liquid handlers, and their ability to capture very high yields and concentrations.  Using 1 mL PhyTip Columns to purify expressed proteins, PhyTip Columns can quickly process 5, 10, 20 mL sample volumes or more by simply repeating the automated steps as many times as necessary.

The Challenges of High Yields with High Throughput

Transient transfection allows for rapid production and screening of high quality drug candidates.  Advances in mammalian cell lines, reagents and other technology are making transient transfection increasingly successful and common. HTP automation of transient transfection is increasingly required to keep up with increased sample load.

However, protein expression volumes are limited when working with transient transfection, which in turn requires greater sample volume to be processed to achieve sufficient target volumes. But, HTP liquid handler are limited in the sample volumes they can process, which can create a bottleneck in the purification process. What is needed is a HTP solution that requires no manual intervention (eg. to clean vacuum manifolds or move to centrifuges), and can be repeated as needed to achieve desired volumes.

PhyTip Columns Provide the HTP Solution

PhyNexus PhyTip Columns are compatible with most major HTP liquid handling instruments.  Moreover, they can accommodate virtually any resin in addition to the standard ProPlus and ProPlus LX resins so commonly used for mammalian protein purification.  Additionally, there is no manual intervention required.  PhyNexus PhyTip Columns can purify up to 1 mL at a time, elute, and repeat until the researcher has accumulated the sufficient target volumes for downstream applications – depending on the robotics used, up to 50 mL.

PhyTip Columns are exceptionally well-suited for transient transfection because the Tip Concentrating Effect produces very high yields and concentrations, even from more dilute samples.  Dual Flow Chromatography’s back-and-forth flow drives a much higher binding reaction than batch or diffusion processes, ensuring the highest amounts of target protein are being captures on each pass.  HTP automation with repeatable steps require no manual intervention, giving researchers the high quality protein they need quickly and easily.