Automation and Separation Process Using PhyTip Columns
Automation and Separation Process Using PhyTip Columns

PhyTip Columns and Automation

What are PhyTip columns?

PhyTip columns are part of a unique protein purification and enrichment system offered by PhyNexus. The PhyTip tips are novel column protein capture devices employing various affinity resins maintained at the base outlet of a pipette tip. PhyTip columns capture the protein of interest by bi-directional flow of sample volume through the column bed. By using back and forth flow, the capture interaction of the protein with the column is driven to completion: capture is complete.

PhyNexus provides the power of chromatography in a pipette tip column in automated liquid handlers for protein and plasmid purification. We provide life science researchers the solution to high through put sample processing in drug discovery, drug screening, assays, and structure work – any process where very high concentrations of very pure product are desired and where it is important that the biological molecule remains undamaged and active. For proteins, PhyTip columns can be used for purification from culture and modifying reactions, expression analysis, purification and immunogenicity screening, protein to protein measurements, CoIP, IP, ChIP, target immobilization, buffer exchange, glycosylation analysis, process development, and biofunctional assays. Through our flexible, custom manufacturing process, PhyNexus can pack most customer resin and chemistries. For plasmids, PhyNexus has developed a proprietary silica resin to purify transient transfection quality plasmids.

Our PhyTip chromatography pipette tip platform enables chromatography for up to 12 samples in parallel and for up to 96 samples in parallel. A wide range of sample sizes can be processed from a broad selection of column bed sizes and chemistries, including affinity, ion exchange, gel filtration, normal phase, and reverse phase.

Can PhyTip columns be used on full automation robotics?

PhyTip pipette columns are compatible with all major robotic systems including Tecan, Agilent, Beckman, Perkin Elmer/Caliper, Dynamic Devices and Hamilton robotic systems. As we understand the goal and the sample provided for the process, we provide the total solution including scripts, robotic set up, installation, etc.

For customers with limited budgets and limited bench space desiring automation, bench top MEA and MMG systems are available. The PhyNexus AutoPlasmid MMG benchtop system is available to produce automated large scale transient transfection quality plasmid.

How are PhyTip columns used?

Virtually any liquid handling robot can use PhyTip columns. The multi-channel head functions simply as a pump. PhyTip columns process is equilibration, sample load, a series of washes and recovery of the sample in final elution step. In addition to fully automated walk-away advantages, the samples can be tracked by analysis of the flow through solutions and washes. The small elution buffer volume maintains the target protein at high concentrations, making the columns especially useful for downstream assays requiring high concentrations.

PhyTip columns are used for leads screening, purification prior to protein analytics, to study protein-protein interactions, and to screen expression and purification conditions.

What are the current competitive technologies used for affinity purification?

Current affinity purification tools include spin columns, gravity columns, magnetic beads, and the FPLC chromatography platform. For small samples, PhyTip tips outperform all of these technologies with higher protein concentrations, higher purity, and higher protein activity. By incorporating Dual Flow Chromatography, the PhyTip sample purification preparation system provides a level of performance unmatched by standard techniques. Purification conditions developed for PhyTip columns are scaleable to preparative and large scale columns. Further information throughout this question and answer section.

How does PhyTip technology compare to FPLC chromatography?

The FPLC chromatography platform performs well and is sometimes considered the “gold standard for protein purification. However, the throughput is limited (samples are run one at- a-time). FPLC set-up and clean-up is time consuming and elution volumes can be large, diluting the final product.

Why would I use PhyTip columns?

PhyTip columns have been optimized to produce the high yield, purity, and activity of sample proteins. They have also been optimized to be able to perform these functions with low volume samples, thus saving valuable resources. Sample work flow is seamlessly integrated with protein express and assays with complete sample and solution tracking. The columns can operate rapidly in parallel operation to perform purification in minimal time, frequently in as little as 15 minutes. The columns are easy to set up and are disposable after use making buffer preparation and cleanup very easy.