ÄKTA avant
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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 chromatography media (resins) 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™ software is used to control automated processes on ÄKTA avant to increase productivity and efficiency. ÄKTA avant offers a complete solution for fast, high-quality protein separations while maintaining reliability and increasing process knowledge. ÄKTA avant offers the following benefits: • ÄKTA avant together with UNICORN software provides a platform for efficient process development • Design of Experiments (DoE), an experimental design tool, provides time and cost savings by capturing more precise information in fewer experiments • 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 software provides intuitive and flexible method creation, system control, and evaluation to simplify your purification task Fig 1. ÄKTA avant is a preparative chromatography system designed for process development, method optimization, and scale-up. System components The system consists of the ÄKTA avant instrument and UNICORN software. The instrument offers easy access to 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 instrument. The wet side of the instrument allows easy interaction with the system, and has a door and pump cover for safer 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 display on the front panel informs you of the current instrument and method state, and the run can quickly be paused or continued from the display. To ensure reliability, the system performs self-diagnostic tests of appropr

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 ÄKTA avant system and sample pumps are based on wellknown technology and robust construction. The pumps deliver reproducible flow rates at both low and high back pressures, allowing short separation times and the use of modern scale-up chromatography media. Figure 2B shows a view of the wet side of ÄKTA avant and the location of the...

• Column valve: Connected to the injection valve and used to select the column 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, minimizing 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 can 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...

Phase Library Method Outline Phase properties pane and Text Instructions pane Column Logbook To increase operational safety, UNICORN software features the Column Logbook. This practical tool keeps track of important column and run data to provide traceability and operational security. Most prepacked columns from GE Healthcare are barcode labeled, and individual columns are identified using a 2-D barcode scanner. The information can also be entered manually into UNICORN (Fig 7). UniTag sticker labels, with preprinted barcodes, are available for other columns (e.g., empty columns). Gradient...

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. 1500 1400 1300 1200 1100 1000 Design of Experiments (DoE) Included as standard for ÄKTA avant systems, UNICORN...

UNICORN method Design input • Definition of factors, factor types, and settings • Definition of objective for creating the design Model evaluation Use of model for prediction and decisions Fig 10. The basic DoE workflow is shown. In the DoE tool, multiple factors are varied simultaneously and the resulting data is used to generate a statistical model. The model is validated and used to produce maps that support decision making. 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 and used...