GE Healthcare Life Sciences Data file 29-0211-96 AB Chromatography systems ÄKTA™ pure ÄKTA pure is a flexible and intuitive chromatography system (Fig 1) for fast purification of proteins, peptides, and nucleic acids from microgram to gram levels of target product. ÄKTA pure is a reliable system where hardware and UNICORN™ software are designed to work together with columns and media to meet any purification challenge. The system supports a wide range of chromatography techniques and meets the automation requirements needed to deliver the highest purity. The system is configurable and can be upgraded at any time with a wide range of options to further increase its capabilities depending on your purification needs. ÄKTA pure is the product of over fifty years of expertise in the development of ÄKTA purification systems. ÄKTA pure offers the following benefits for you: • Modular system design with a large range of options to allow flexibility in purification of proteins and peptides • Customizable system that is easy to upgrade as your research needs develop • Reliable system with components and integrated features based on the proven design of ÄKTA avant • UNICORN 6 software provides simple, intuitive, and flexible preprogrammed method templates and total system control to simplify your job • Predefined method settings for all GE Healthcare lab-scale chromatography columns System overview ÄKTA pure chromatography system is a highly versatile, modular system with a number of design features to facilitate reliable purification. The system consists of the ÄKTA pure instrument and UNICORN 6 control software. The system is modular in design with all valves, monitors, and columns mounted on the forward facing wet side of the system, to allow easy interaction with the instrument modules (Fig 2). Additional components such as valves, monitors, and sensors from the wide range of optional modules can be easily added Fig 1. ÄKTA pure is a flexible chromatography system for the reliable purification of proteins, peptides, and nucleic acids at laboratory scale. to the available positions. Multiple rails for attachment of column holders and equipment are located at the front and on the side of the instrument. A buffer tray on the top of the instrument provides a large storage area for vessels and bottles. The instrument control panel shows the system state and allows the possibility to interact with the run (pause/ continue) at the touch of a button. The system weighs only 48 kg in basic configuration and 53 kg when fully equipped with options. The relatively low weight enables easier placement in the lab and the system dimensions allow it to fit conveniently into a standard cold cabinet for work with labile samples.

Column valve V9-Cs with built-in bypass and reverseflow functionality UV monitor U9-L for detection at 280 nm Rails for attachment of column holders and accessories (also on the left-hand side) Tubing organizer Column valve V9-Cs with built-in bypass and reverseflow functionality Verify gradients with Conductivity monitor UV monitor U9-L for detection at 280 nm Round fraction collector F9-R for collection in various tube types Tubing organizer Inlet valve V9-IAB allows selection between two A and two B positions in a single valve Verify gradients with Conductivity monitor High-performance system...

Regardless of configuration, ÄKTA pure always comes with two high-performance system pumps, system pressure monitor for column protection, mixer, injection valve, and UV monitor. ÄKTA pure has a wide range of optional modules to allow a large number of possibilities. The system flow path is designed to minimize band-broadening effects, and all wetted materials used in the flow path are biocompatible and resistant to commonly used solvents. The instrument front is designed with empty module positions where optional valves and monitors can be mounted to enable a flexible configuration of the flow...

Flow rate: System: 150 Superdex 200 10/300 GL Molecular weight standards for gel filtration 100 µl PBS (10 mM sodium phosphate, 140 mM NaCl, 2.7 mM KCl, pH 7.4) 0.5 ml/min ÄKTA pure 5000 5 A280 nm A214 nm A340 nm 4000 2 3 4 Column: Sample: Sample volume: Eluent: The optional sample inlet valve is intended to be used with the sample pump. It allows fast, automatic loading of up to 7 different samples. The integrated air sensor ensures safe and complete sample application without the need to preprogram the sample volume. The valve has seven sample inlet positions plus a dedicated buffer inlet for...

The A420 signal specifically monitors the target protein. The columns giving the sharpest and most symmetrical peaks at A420, as well as the highest possible purity, were selected for subsequent optimization and scale-up experiments. HiTrap Phenyl FF (high sub) 1 ml and HiTrap Butyl FF 1 ml gave the most promising results under the conditions used, and HiTrap Phenyl FF (high sub) 1 ml was selected for further optimization in this case. Conductivity (mS/cm) Conductivity (mS/cm) Conductivity (mS/cm) Conductivity (mS/cm) HiTrap Phenyl FF (high sub) 1 ml The flexibility of Column selection valve...

Outlet valves Two different valve options are available to direct the flow to the fraction collector, waste, or other outlet ports. Outlet control valve V9-Os allows connection of one or two fraction collectors. If only one is connected, the other port can be used for outlet fractionation, for example to collect flowthrough. Outlet fractionation valve V9-O enables connection of up to two fraction collectors and up to 10 available outlets allow collection of large fractions. The cassettes can also be used for convenient storage of fractions or holders for sample tubes and are easy to handle and...

UNICORN 6 control software Phase Library Method Outline Phase properties pane and Text Instructions pane UNICORN control software is used on all ÄKTA systems and can be used from lab scale up to full-scale production. UNICORN contains the tools needed for beginners and experienced users to perform all types of chromatography, from setting up and running a method to evaluating the data. Operation of ÄKTA pure is controlled by UNICORN 6 system control software, which provides built-in knowledge for planning, controlling, and analyzing the system and chromatography results. For greater convenience,...