Affinity chomatography is a routinely used technique for purifying or enriching a protein or molecule of interest through its specific binding affinity. The target protein adheres to a particular ligand that has been immobilized on a solid support (usually beaded agarose resin). This process produces high selectivity, resolution, and capacity for the protein of interest. Traditionally, affinity chromatography is performed in column format. Where the sample is applied and eluted by gravity flow through a packed resin bed of one to several milliliters. However, gravity flow columns are time consuming and require constant attention while supernatant filters through the column and resin. This process can be accelerated with G-Biosciences’ FastPure™ Spin Columns (Mini and Midi) specifically designed for simple and efficient small scale protein purifications. FastPure™ Spin Columns combine the effectiveness of gel filtration, and the speed of centrifugation for quick and reliable protein purification.
While the traditional gravity column format is suitable for applications that involve large quantities of immobilized ligand and purified product, a smaller batch format is more suitable when the sample volume is small and only microgram amounts of purified protein are required for subsequent analysis (e.g., by gel electrophoresis and Western blotting detection). They come in two sizes, Mini and Midi, which conveniently fit into common centrifuge tubes for quick and easy wash and elution steps. The Mini spin columns fit into standard 2 ml microcentrifuge tubes (maximum volume: 600 µl) and the Midi spin columns fit all 50 ml conical centrifuge tubes (maximum volume: 20 ml) to be used with standard bench centrifuges containing a swing bucket rotor.
G-Biosciences’ FastPure™ Spin Columns have a unique, proprietary membrane that retains resin and supernatant during incubation and mixing, but that opens up when the columns are centrifuged to allow only the supernatant to pass through. The retained resin can be washed and proteins of interest eluted simply by adding appropriate buffer and centrifuging.
These versatile spin columns are compatible with all chromatography resins (protein A/protein G, IMAC, glutathione, etc). Simply add the desired affinity resin and supernatant or lysate to the column then incubate (batch mix) your sample containing the protein of interest with the desired resin in buffer conditions that facilitate binding. After incubation, centrifuge the column to remove the unbound supernatant and then wash with additional buffer to wash away nonbound components of the sample from the resin. Finally, the molecule of interest is recovered (eluted) from the resin using an altered buffer condition that disrupts the ligand-molecule binding.
G-Biosciences’ FastPure™ Spin Columns allow you to wash and elute your purified and concentrated protein all in one chamber. These innovative spin columns incorporate a specialized low protein-binding ultra high density polyethylene membrane that retains the sample and resin in the column and prevents leakage. When the column is centrifuged (Mini: 12-14,000 x g for 1 min, Midi: 750-1,500 x g for 5 min) the pores of the membrane dilate (0.1-0.2 µm) and allow the sample to be purified and collected for subsequent downstream analyses. There is no resin carry-over, no messy transfer of samples or resin slurries, and negligible hold-up volume. These columns are self-contained and easy to dispose of, which makes them ideal for handling biohazardous and infectious samples.
G-Biosciences’ FastPure™ Spin Columns effectively produce highly concentrated purified protein. These columns save valuable time by allowing you to efficiently process multiple samples in parallel. The innovative membrane prevents leakage and allows you to perform fast and effective incubation, wash and elution steps in one self-contained, disposable column. Easily collect your purified or concentrated protein in any standard 2 ml or 50 ml centrifuge tube. These versatile spin columns are compatible with all affinity chromatography resins, reagents and buffers (G-Biosciences’ Purification and Chromatography products).