GE Healthcare Data file 28-9496-70 AA Chromatography systems ÄKTAmicro™ ÄKTAmicro chromatography system (Fig 1) is designed for microscale liquid chromatography applications. Reliable, high-resolution purification of target molecules is achieved from extremely small amounts of starting material and from molecules as diverse as intact proteins, protein complexes, and nucleotides. Moreover, ÄKTAmicro is an excellent choice for characterization of target molecules and for rapid purity/ recovery analysis in method development and drug discovery applications. The system is built on the proven ÄKTA™ design platform and is controlled by UNICORN™ software. ÄKTAmicro chromatography system offers: • High-performance purification and characterization using microbore to analytical scale columns, all controlled by UNICORN software • High sample recovery and stability as the complete flowpath is manufactured from inert and biocompatible materials • Optimized resolution by connection of columns directly to the UV cell and by minimized dead volumes in the tubing, valve, and flow cells • Novel pump design for flexible flow and pressure range enabling high- as well as low-pressure separations • Fast and reproducible purity analysis by gel filtration in method development and drug target characterization • Absolute characterization of biomolecules in solution and on-line molecular weight analysis by the addition of light scattering equipment Fig 1. ÄKTAmicro chromatography system allows purification of extremely small sample volumes and is an excellent tool for rapid purity analysis in method development and drug target characterization. Fraction Collector Frac-950, ÄKTAmicro offers microliterscale fraction volumes and excellent chromatographic performance. Autosampler A-905 provides reliable and reproducible sample injection and unattended, multiple sample runs. Table 1. Column diameter scales suitable for use with ÄKTAmicro system Modular design based on the proven ÄKTA design platform Purification scale Microbore ÄKTAmicro is modular by design and is comprised of three main components: Pump P-905, Monitor pH/C-900, and Monitor UV-900. Autosampler A-905 and Fraction Collector Frac-950 are optional components. The system is based on the proven ÄKTA chromatography platform and is optimized for microbore-, narrowbore-, and analytical-scale purification of proteins and peptides (Table 1). In combination with Analytical scale

Accurate and reproducible flow rates and gradients ÄKTAmicro generates highly accurate and consistent flow rates and gradients over the entire working range. Pump P-905 is incorporated in ÄKTAmicro and overcomes many of the problems associated with traditional high-performance pumps at low flow rates. The flexible flow and pressure range even enables low-pressure separations, making the system compatible with soft agarose-based media for fast, reproducible gel filtration. This low-pulsation pump is equipped with four pump heads in two binary modules, which allows for binary gradients with high...

Flow configuration The standard analytical flow configuration of ÄKTAmicro is shown in Figure 4. This flow configuration enables chromatography with 1 mm i.d. columns or greater. Direct connection of the column to the UV- and conductivity monitors minimizes dead volumes, which ensures accurate gradient reproducibility and improves purification resolution. even for partial loopfill injections down to 5 μl. For small sample volumes, the microliter pick-up procedure allows injection with no sample loss while thorough wash routines efficiently eliminate cross-contamination. Accurate on-line gradient...

AD-900 analog/digital converter If external instruments are to be connected to UNICORN, the AD-900 analog/digital converter is needed. AD-900 has an analog input for monitoring signals received as voltage outputs from external sources, such as fluorescence detectors, radioactivity monitors, refractive index monitors, or diode arrays. Valve INV-917 This seven-port valve can be used as an additional valve together with the optional software strategy for customized applications. High-resolution columns for reliable target molecule purification Fig 8. UNICORN provides fast and user-friendly interaction...

Reliable results, whatever the application Micropurification There are three main reasons to perform purification in microscale: 1) only small amounts of sample are available, 2) low abundance of target protein in the sample, 3) to keep consumption of reagent and sample down to minimize cost. Figure 10 shows the micropurification of PEP-19 (Purkinjecell protein 4; PCP 4, Mr 6700), a brain protein involved in calmodulin-dependent signal transduction. To demonstrate the suitability for micropreparative protein purification using ÄKTAmicro, a protein extract from mouse brain tissue was prepared...

Column: Sample: Superdex 200 PC 3.2/30 Monoclonal IgG4, fractions from purification step using MabSelect SuRe affinity chromatography medium Elution buffer: Phosphate Buffered Saline (PBS) Flow rate: 0.04 ml/min Fraction size: 40 µl Detection: UV 280 nm System: ÄKTAmicro Specifications System specifications 2 to 12 4°C to 40°C All commonly used chromatographic solvents 0.15 mm i.d. Peak tubing dimension Conductivity range Solvent compatibility Pressure range Temperature range Flow rate range Flow rate range Fig 11. Purity analysis of IgG4 on Superdex 200 PC 3.2/30 gel filtration column and ÄKTAmicro...

Conductivity unit Deviation from theoretical conductivity Maximum ± 2% of full-scale calibration range or ± 10 μS/cm, whichever is greater in range Rack B (240 × 12 mm tubes), supports Eppendorf tubes Rack C (4 × microtiter plates and 8 × 30 mm tubes) Maximum ± 0.5% of full-scale calibration range Conductivity range Flow cell Delivered with Frac-950 as standard Maximum flow rate Maximum pressure Ordering information Internal volume System and components ÄKTAmicro system Autosampler A-905 Sample capacity Plate with 48 wells for standard, 1.5 ml vials includes ÄKTAmicro, software, and installation...