AKTAprime is an entry-level, low-pressure (1 MPa max with flow rate of 0.1-50 ml/min) chromatographic system from the GE Healthcare/Amersham Biosciences line-up, designed primarily for protein purification. It is a small footprint, all-in-one system featuring a built-in rotary fraction collector, in-line UV monitor (selectable 254 and 280 nm with options for 214, 313, 405, 436, and 546 nm), a conductivity meter (0.001-999.9 mS/cm), and gradient maker/mixer. There are only a few options including a pH monitor, an assortment of mixing chambers and varying column holders, offering little flexibility over and above the basic design.
The system is operated from the on-board chip through a simple, albeit somewhat counterintuitive, front-panel keyboard and can be used with either a dual channel recorder (Rec-112) or with the PrimeView software package. It would be preferable to use the recorder in a cold room (due to electrical hazards associated with computer use in this environment). It is important to note that the recorder only captures two signals (e.g. UV absorbance and conductivity) in addition to marking “events” (such as fraction collector advancing). PrimeView is adequate but somewhat cumbersome and dated. This software package simultaneously displays the readings of run parameters and the event log, while enabling the user to perform limited editing of the recorded run (can be imported into graphics software or printed out).
The system can be operated in a variety of ways including manual operation (real-time), pre-programmed protocols or custom/user-defined protocols. Pre-programmed protocols (“Application Templates”) include such popular and easily reproducible techniques as His6-tag/GST-tag affinity purifications and purification of monoclonal antibodies, all utilizing pre-packed media. The design of these templates is straightforward enough to enable a non-biochemical lab to successfully employ these techniques (e.g. for antigen purification or antibodies clean-up in an immunological lab).
AKTAprime can utilize an appreciable array of Amersham/GE Healthcare pre-packed media, namely HiTrap, HiLoad, HiPrep, as well as resource columns for affinity, gel-filtration, ion-exchange and hydrophobic interaction chromatography.Unfortunately, none of the Mono media can be used with this system. In addition, some low pressure limit/high flow columns (e.g. gel-filtration) require changing the standard set-up of the system to PEEK tubing with larger inner diameter in order to function properly.
Our lab has used 2 AKTAprimes, one with the recorder and another with a laptop running PrimeView, for over 2 years and our impression has been rather positive. The system works well right out of the box and although not as versatile as FPLC, performs rather impressively, especially with the Resource media. By introducing appropriate affinity (metal, heparin, etc.) steps together with ion-exchange (Resource Q, SP), we are able to purify several proteins for NMR and X-ray crystallography solely on AKTAprime. The overall reliability of the AKTAprime system has been rather good, with only a handful of breakdowns usually involving sensors for the injection and buffer valves, but the pumps, mixers, and the optics have performed very admirably.
Vladimir Svetlov, PhD
Artsimovitch Lab
Department of Microbiology
Ohio State University