Protein Purification

Better, Easier, Faster – Protein Purification with PhyTip Columns

PhyNexus proprietary PhyTip Columns deliver superior protein purification results for virtually any application in a format that is easier, faster and more reliable than traditional columns or batch binding processes. Protein purification using PhyTip Columns with Dual Flow Chromatography techniques drives the sample binding reaction to equilibrium, resulting in superior yields and concentrations, and because the process of Dual Flow Chromatography is gentle, it also delivers superior samples in terms of purity and activity.  And every PhyTip Column is designed to be compatible with high throughput or bench top automation.

The Challenges of Small Scale Protein Purification

The ultimate goal of small scale protein purification has always been to combine high throughput processes with excellent sample results. Unfortunately, combining these two objectives in one process has proven challenging, and most advances in small scale sample preparation have many drawbacks. FPLC, while effective, has very low throughput and significant hands-on time.  Spin columns, while effective, are low throughput, require manual intervention for centrifugation, suffer from lower reproducibility and can be harsh on fragile protein complexes. Magnetic bead approaches, while capable of high throughput, suffer from low capacity which results in lower yields and concentrations and magnetic beads can be quite costly.

Additionally, none of these other processes allow for extremely fine and precise control of the chemical environment and reaction mechanics, such that researchers and fine tune reactions.

PhyNexus Delivers High Yields and High Throughput (and more) at Microscale

PhyNexus PhyTip columns combine high throughput automation with superior sample results. PhyTip columns utilize active transport of sample across a solid immobilized between two hydrophilic frits in the tip of the column. Using Dual Flow Chromatography, the sample is gently drawn up from the plate well into the tip and resin bed. Then, the sample is gently expelled back down into the plate well. This gentle back and forth flow is repeated an optimal number of times – usually three to four cycles – driving the binding reaction to equilibrium, a result that is not possible with linear flow processes or batch binding processes.

PhyTip columns offer an array of other advantages for protein purification. Because they are small, only small molar excess of expensive binding reagents is required to bind the target molecule to the resin. And because PhyTip columns are small, only small total amounts of reagents are required. The volume of sample needed to load the column is also small, so high concentration of loading molecule can be used, driving the loading reaction to completion more quickly and with less reagent.

As if all that wasn’t good enough, PhyTip columns are purposefully designed to be compatible with high throughput automation. Automation delivers the advantages of high throughput, increased productivity, less experimental variability, reproducible results and greater confidence. PhyTip columns are compatible with virtually all major liquid handling instruments, as well as the PhyNexus 12-channel bench top MEA 2.