ÄKTA avant CHROMATOGRAPHY SYSTEMS
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GE Healthcare Life Sciences Data file 28-9573-45 AE Chromatography systems ÄKTA™ avant ÄKTA avant is a preparative chromatography system intended for method and process development (Fig 1). Available in two versions, ÄKTA avant systems have different flow rate/pressure specifications but share the same hardware setup. With flow rates up to 25 ml/min, ÄKTA avant 25 system is designed for media screening and method optimization. ÄKTA avant 150 system, with flow rates up to 150 ml/min, is designed for scaling up to larger columns. UNICORN™ 6 control software has been specially developed for ÄKTA avant to increase productivity and efficiency. ÄKTA avant offers a complete solution for fast, high-quality protein separations while maintaining flexibility and reliability. Fig 1. ÄKTA avant is a preparative chromatography system designed for process development, method optimization, and scale-up. Key features and benefits: • Design of Experiments (DoE), an experimental design tool integrated in UNICORN 6, provides time and cost savings by capturing more precise information in fewer experiments • Column recognition and run data history of individual columns provides traceability and operational security • Integrated fraction collector with cooling functionality protects purified samples • Automatic on-line buffer preparation using BufferPro reduces the time required for buffer blending and manual titration, increasing your productivity • UNICORN 6 provides easy protocol transfer during scaleup and an improved visual interface for intuitive and flexible method editing System components The system consists of the ÄKTA avant instrument and UNICORN 6 control software. The instrument offers easy access to the working areas using a swivel foot (Fig 2A). ÄKTA avant has a modular design, with all valves, monitors, and columns mounted on the wet side of the system. The wet side of the instrument allows easy interaction with the system, and has a door and pump cover for safe handling during runs (Fig 2A and 2B). A buffer tray on top of the instrument provides a large storage area for vessels and bottles. On the front side of the instrument, ÄKTA avant has a built-in, cooled fraction collector that provides secure sample storage. An interactive instrument display on the front panel informs the user of the current instrument and method state, and the chromatographic run can quickly be paused or continued from the display. To ensure reliability, the system performs self-diagnostic tests of appropriate settings at start-up.

Large buffer storage area Interactive touch display Door and pump cover protects flow path Integrated fraction collector Storage box Swivel foot for easy access to all sides Sample placement Wet side with modules B) UV monitor Conductivity monitor Sample pump • System pump: Provides continuous and accurate flow rates, to give reproducible isocratic or gradient elutions. This pump consists of two pairs of pump heads, which deliver synchronized, low-pulsation flow to the mixer. For ÄKTA avant 25, the maximum operating pressure is 20 MPa and the maximum working flow rate is 25 ml/min. For ÄKTA...

• Column valve: Connected to the injection valve and used to select the column that is to be used. Up to five columns can be connected to the column valve for automatic column scouting. The column valve has an integrated column bypass function, eliminating the risk of overpressure on the columns, for example, during system and pump wash procedures. To increase the yield and performance when using absorption techniques, the column valve also has a built-in reverse flow functionality. Reversed flow may be used during column cleaning or when eluting a sample to give sharper bands and a more...

• UV monitor: Provides real-time measurements of absorbance over the UV and visible range of 190 to 700 nm. To visualize protein separation at different wavelengths, the UV monitor has a flip-mode that allows monitoring of up to three wavelengths simultaneously. For optimum performance when purifying samples with different protein concentrations, there are three flow cell path lengths (0.5, 2 [default], and 10 mm). The optimal flow cell design, together with advanced fiber optics, provides a high signal-to-noise ratio without causing any local heating of the UV flow cell (especially...

UNICORN 6 control software UNICORN 6 control software, specially developed for ÄKTA avant, gives you real-time control of your chromatography system. UNICORN 6 consists of four modules: Administration, Method Editor, System Control, and Evaluation. This section describes some of the valuable tools included in UNICORN 6 for increasing operational security, efficiency, and productivity. Method Editor The Method Editor contains all the instructions used for controlling the chromatographic run. In UNICORN 6, the improved Method Editor has a user friendly graphical interface for easy viewing and...

BufferPro Automatic buffer preparation with BufferPro increases productivity by minimizing the need for manual preparation of buffers. BufferPro can be used for automatic pH scouting to find optimal buffer compositions. For screening, a broad pH range is scanned by testing large pH steps. For further optimization and to test robustness, smaller pH steps are tested (Fig 8). BufferPro can be used at flow rates up to 25 ml/min on ÄKTA avant 25 and up to 40 ml/min on ÄKTA avant 150. BufferPro eliminates time-consuming buffer preparation and manual titration for experiments requiring pH changes....

The resulting data or “responses” (e.g., yield and purity) are used to create a statistical model (Fig 10). This model is automatically generated in UNICORN 6 and used to predict the response to changing a factor, and to produce maps (response contour plots) that support decision making. Because DoE estimates variability and noise as well as interaction effects between different factors, more precise information is obtained and better maps are generated, which leads to better decision making. Table 2. Summary of the method development steps in a two-step MAb purification using ÄKTA avant 25...