Short on protein chromatography time?
By GE Healthcare
Wednesday, 11 December, 2019
Have you ever considered how a resin bead in your chromatography column can affect your ion exchange (IEX) chromatography run time?
In a recent webinar, we shared some tips for improving protein purity and recovery while performing IEX chromatography; here, we focus on your time. Today’s protein researcher is pressed to deliver results faster than ever before. The less time spent on purification, the more time you will have to focus on studying your protein.
Ion exchange chromatography can be used in the first steps of protein purification for both native proteins and untagged recombinant proteins. While protein purification has become almost routine, new technology and materials always create opportunity to look for improvements. That is the case when choosing a Capto ImpRes resin for use with viscous samples.
Speeding up run time for viscous samples
Viscous samples, such as E. coli extract, can increase back pressure on the column and force you to reduce the flow rate, resulting in longer load times. Utilising a robust, modern and rigid resin — such as Capto Q ImpRes — reduces sample run time significantly by allowing high flow rate with low column backpressure, as seen in the example below.
HiTrap Capto ImpRes contains Capto ImpRes high-flow agarose resin, which has much better pressure/flow properties than Sepharose High Performance resin. (Capto ImpRes resin has maximum flow rate of 300 cm/h vs 150 cm/h for Sepharose High Performance.) This allows more flexibility — even in the HiTrap format — for viscous samples such as E. coli lysate, and allows them to be loaded at higher flow rates in a HiTrap Capto ImpRes than a HiTrap HP column. In this example, the IEX total run time is 30% shorter with HiTrap Capto Q ImpRes than a HiTrap Q HP, with an otherwise similar performing column.
HiTrap Capto ImpRes: a versatile tool
HiTrap Capto ImpRes columns can be used for viscous samples as well as for other high-resolution IEX chromatography procedures with a broad range of samples. Since the resolution is similar to Sepharose High Performance, Capto ImpRes can be used in these applications as well, and there is no need to have both columns on hand.
Capto ImpRes resin has been used in diverse applications, including:
- analysing the influence of a mitochondrial protein in mammalian cytosolic Fe-S repair
- researching the mode of action of antimalarial drug primaquine
HiTrap Capto ImpRes can serve as a universal column for routine work in a research lab. Use HiTrap Capto Q ImpRes for anion exchange chromatography, and HiTrap Capto SP ImpRes for cation exchange chromatography.
To learn more about our new Ion Exchange chromatography resins, download our newly revised handbook here.
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