Chromatographic equipment and media
The chromatographic methods employed at the ISPC are tailored to the physical and chemical properties of the individual protein targeted. Commonly used chromatography techniques include: size exclusion chromatography (SEC; gel filtration), ion exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), and affinity chromatography using resins bearing Ni+2, GST, streptactin, Protein A, amylose, FLAG, or chitin, as well as affinity resins bearing ligands specific for the proteins of interest. These resins may be utilized as slurries or in prepacked columns.
The center is equipped with four AKTA (GE Healthcare) FPLC systems, all of which are operated in refrigerators, thus permitting execution of the entire purification process at 4°C. All four systems are equipped with UNICORN software to control and record the process, with UV detectors, and with fraction collectors.
The AKTA explorer at the ISPC is equipped with a sample pump P-960 and a
Frac-950 fraction collector, as well as a 3D plus kit. It allows automated multi-step purification of tagged proteins. The AKTA 3D plus kit is intended specifically to permit automated preparative purification of soluble His- and GST-tagged target proteins. Together with the AKTA explorer the combined hardware and software provide a choice of five automated purification protocols that contain up to three chromatographic steps. The system allows two-step purification of up to six samples in one automated run.
AKTApurifier & AKTApure
AKTA pure & AKTApurifier are flexible and intuitive chromatography systems for fast purification of proteins. They are both modular systems designed with a large range of options to allow flexibility and reliable protein purification. Both systems employ the UNICORN software.
Autosampler A-905 for AKTApurifier
The AKTApuifier is equipped with an A-900 autosampler that offers a variety of alternatives for sample injection with high reproducibility. The autosampler allows injection of even small analytical samples without any losses. Moreover, it is controlled by UNICORN software as an integral part of the AKTApurifier. Thus allowing multiple injections in an automated manner.
The QuixStand benchtop system is a laboratory-scale cross-flow hollow-finer system designed for fast, efficient concentration and/or diafiltration of biological solutions. It is capable of rapidly processing solution volumes from as little as 500 ml up to 10 l. The ISPC has two compatible hollow-fiber ultrafiltration cartridges (3.5 & 10 kDa MWCO). The system consists of a cartridge support stand, inlet and outlet, pressure gauges and a 400 ml reservoir. The QuixStand system provides an efficient method to concentrate large volumes of media, and to perform buffer exchange. It is commonly used prior to purification of proteins from culture media.
Benchtop cell disrupter
This high pressure cell disruptor is suitable for thedisruption of any type of cell. Its main advantages are that it is easy to use, and maintains the cells and lysate at 4°C throughout the procedure.