PhyTip® Ion Exchange columns offer the unique ability to screen different wash and elution buffers and a number of resins in a single experiment. Once ion exchange conditions have been optimized, they are directly transferable to larger manufacturing scale columns. Chromatographic separation is performed in a pipette tip column using back and forth flow to drive binding and release steps to equilibrium. Using the MEA 2 Personal Purification System enables 12 samples at a time to be processed. This can be increased to 96 samples with the use of robotic liquid handling systems. Four standard ion exchange resins are available in PhyTip columns (strong anion, weak anion, strong cation, weak cation). For resin selection, the Ion Exchange Resin Screening Toolkit is a box of 96 tips containing 24 columns of each resin.

One advantage to ion exchange is the buffer conditions used for capture washing and elution are generally compatible with the proteins so that high activity can be retained in the final product. Moreover, ion exchange chromatography may still be used for final purification and polishing. Using ion exchange as an orthogonal chemistry to affinity chromatography can result in very pure proteins.

There are two basic types of ion-exchangers: cation exchangers and anion exchangers. Cation exchange separations are performed at pH’s between 4 – 7 and anion exchange separations are generally done at pH’s between 5 – 8.

Within anion exchange there two types: strong base anion exchange and weak base anion exchange. The same is true for cation exchange: strong acid cation exchange and weak acid cation exchange.

Choosing the most effective ion exchanger and ion exchange conditions ion exchange purification may be difficult to determine. The PhyNexus PhyTip column technology based on pipette tip columns can easily process samples in parallel making it effective to screen dozens to hundreds of ion exchange within a very short time.

Ion Exchange columns with our proprietary Dual Flow Chromatography provide complete antibody binding and high final concentrations. PhyTip columns are compatible with most 8, 12, and 96-channel automation liquid handling robots including Agilent TechnologiesBeckman CoulterDynamic DevicesPerkin ElmerHamilton, and Tecan.

Description

PhyNexus’ family of PhyTip products includes ion exchange. In traditional ion exchange chromatography, samples, wash buffers and gradient elution buffers pass through packed columns in one direction. The PhyTip column is able to simulate this chromatographic separation in a small, pipette tip column by driving the binding and release steps to equilibrium. This is not achieved by using high back-pressure, large columns, but instead by PhyNexus’ patented back and-forth flow.

  • Develop ion exchange purification methods in a fast, efficient manner
  • Automation compatible on the MEA Personal Purification System or other robotic systems
  • Take advantage of parallel processing to screen resins
  • Applications include conditions optimization, imidazole removal after His-tagged protein purification and high-throughput replacement of finishing columns
Documents
Compatibility

Ion Exchange PhyTip Columns are available for these robotic systems.

Citations

Posters


Screening for Chromatographic Success and Failure on Small Scale
W. Gillette (NCI/SAIC-Frederick)

Space is the Place: Using DOE and Microscale Techniques to Define Process Boundaries
M. Wenger (Merck & Co.)

Development of Small-Scale, High Throughput, Chromatographic Protein Purification
K. Valente1, M. Wenger2, J. Shanter1, S. Sagar1, J. Konz1 (1Merck Biopurification Development, 2Merck Bioprocess and Bioanalytical Research)

A flexible platform for automated high-performance protein purification using micro scale separation technology
J. Lambert, M. Anderson, D. Gjerde, L. Nguyen (PhyNexus)

Innovative Solutions for Automated Protein and Nucleic Acid Purification
Lee Hoang, PhyNexus, Inc., PhyNexus Webinar

Automated, High Throughput Protein Purification and Sample Prep Using PhyTip Columns for Therapeutic Leads Screening and Process Development
Lee Hoang, PhD., PhyNexus, Inc., The 5th Annual Proteins Congress 2012, London

Optimization of Protein Purification Using Small-Scale Separation Columns
Chris Suh, PhD., PhyNexus, Inc., Protein Therapeutics Discovery and Development

PhyNexus Users Group Symposium 2014
Douglas Gjerde, PhyNexus
Lee Hoang, PhyNexus
Tyson Bowen PacBio

Select your resin, robotic system, tip and resin volume and see the results in the table below.

Ion Exchange PhyTip Columns