Fraction Collectors: Frac-950 and Frac-920

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GE Healthcare Data file 18-1153-57 AE Fraction collectors Fraction Collectors: Frac-950 and Frac-920 Introduction Fraction collectors Frac-950 and Frac-920 are automated for reliable and flexible collection of fractions based on time or volume as part of an ÄKTA design chromatography system. Frac-950 (Fig 1) has advanced functionality for both standard and preparative mode that allows large volume fraction collection. Frac-950 can also easily be equipped with an optional Microfraction Collection Kit, enabeling reproducible collection of volumes down to 40 µl. Frac-920 (Fig 2) has a carousel design, small footprint, and can be used with laboratory systems or as a stand-alone instrument. Reliable and easy-to-use Robust design No sample loss Simple programming Cold room compatible Fig 1. Fraction collector Frac-950. Collect in the vessel you need Wide range of standard collection vessels Preparative mode for large volume fractions with Frac-950 Collect the fractions you want Peak fractionation based on slope or threshold Time- or volume-based collection Microfraction Collection Kit, for reproducible collection of fraction volumes down to 40 µl Fig 2. Fraction collector Frac-920.

Frac-950 for advanced flexible fraction collection Fractions can be collected from practically any size of labscale chromatography column, from 96-well microplates, standard 12, 18 or 30 mm tubes or in 250 ml bottles. Even larger vessels can be used at flow rates up to 100 ml/min. Spillage and sample loss between vessels at low and high flow rates is prevented by drop synchronization or by the built-in accumulator. Peak detection is based on slope or threshold detection, allowing fractionation to be controlled with the minimum of peak overlap and dilution of significant fractions. Frac-950 is controlled simply and directly by UNICORN as an integral part of your ÄKTAexplorer, ÄKTApurifier, ÄKTAfplc, or ÄKTAmicro system. Collect in the vessel you need Collecting fractions in the right kind of vessel helps optimize workflows, whether checking the results of cloning experiments or preparing large amounts of material for a routine application. In the standard mode, fractions can be collected in 96-well microplates (up to four) and 12 mm, 18 mm or 30 mm standard tubes (Fig 4). To avoid collecting unnecessary numbers of small fractions, standard racks A and C have extra positions where larger fractions from wash steps can be collected in 30 mm tubes. Large volumes are managed in preparative mode that allows fraction collection in eighty 30 mm tubes, twenty 250 ml bottles or in thirty funnels (Fig 5) connected to practically any vessel. Robust design with few moving parts Mechanical reliability is essential in any fraction collector so the smaller the number of moving parts the better. In its standard mode, Frac-950 uses a stationary collection head and two simple rotational movements (Fig 3) to position the collection vessel accurately and quickly to collect the next fraction. Large volume fractions are collected in the preparative mode by moving the collection head with the same two motions. Rack A: 120 18 mm tubes 8 30 mm tubes Rack B: 240 12 mm tubes Rack C: 4 microplates (96 fractions per plate) 8 30 mm tubes Rack D: 45 30 mm tubes Rack E: 45 30 mm tubes Rack F: 20 250 ml bottles Fig 4. Available racks for Frac-950. Rack E and F require a Prep Mode Conversion Kit. Fig 3. The Frac-950 positioning system. Using Rack C, each well in four microplates and each of eight standard 30 mm tubes can be positioned precisely and reliably under the stationary collection head. Two independent ways to prevent sample loss Valuable materials must be collected without spillage or loss between collection vessels. Frac-950 achieves this both at low flow rates and at high rates up to 100 ml/min. At low flow rates, fraction changes are synchronized to occur between drops. Since the collection head is stationary in the standard operating mode there is no risk of hanging drops being flicked off. At high flow rates, liquid flow can be diverted to an accumulator during the short time it takes to move to the next collection position. The diverted liquid is then dispensed and collected securely and completely. Fig 5. Frac-950. The preparative mode also accommodates up to thirty very large fractions. 2 18-1153-57 AE

Frac-920 for robust fraction collection Fraction collector Frac-920 has functionality and design that makes it ideal for lab-scale chromatography. The robust design with a minimized footprint also saves bench space. The fraction collector is programmable and easy to use in connection with UNICORN and ÄKTA design chromatography systems or as stand-alone instrument. Fixed volume and time fractionation as well as automatic peak fractionation are possible. Peak fractionation is based on peak detection using slope or level sensing and event marks clearly identify the fractions and peaks. Frac-920 comes with three tube racks to allow collection of up to 95 fractions in 10-18 mm diameter tubes or up to 175 fractions in 12 mm diameter tubes and up to 40 fractions in 30 mm tubes. An optional flow diversion valve SV-923 allows flow diversion to waste. Simple programming with UNICORN Both Frac-950 and Frac-920 can be run in many different ways to collect the fractions you need for your further work. Operation as an integral part of your ÄKTA design system is simple and direct using UNICORN 3.2 or higher (Fig 6). For prep mode Frac-950 requires UNICORN 4.0 or higher. Select rack type Select fractionation order If the same small fraction size is used throughout the separation, then there will be many fractions at the beginning containing nothing of interest, simply making work. If the fraction size is made large to reduce the number of tubes to be assayed, then proteins separated on the column will be re-mixed in the collected fractions. By analyzing the monitor signal, UNICORN can determine just when the separated proteins are eluted from the column and collect fractions to match. As can be seen from Figure 7, cross-contamination of proteins can be significantly reduced by monitoring the peak slope. When a shoulder on a peak or a valley between two incompletely resolved peaks is detected, UNICORN can instruct the fraction collectors to change fractions at the best time to preserve resolution. Between well-separated peaks or at the beginning or end of a run, fractions with little or no protein can be diverted to waste or collected as a few large fractions as a safeguard (racks A and C have extra positions for 30 mm tubes that are ideal for this purpose). 15.0 10.0 mau UV Cond Fractions Minimizes crosscontamination ms/cm 40.0 30.0 20.0 5.0 Minimizes dilution 10.0 Select tube type and size Select fractionation size Select fractionation order Select tube type and size 0.0 0.0 5.0 10.0 15.0 20.0 ml Fig 7. Automatic peak fractionation can help reduce peak overlap and peak dilution. Collection of small sample volumes Micropreparative purification has rigid requirements when working with accurate and reproducible collection of small sample volumes. The optional Microfraction Collection Kit (Fig 8) allows reliable collection of microliter fraction volumes as low as 40 μl directly into microplates (the recommended volume range is from 500 μl/min and below). Fig 6. UNICORN defines your fractionation scheme to suit your exact requirements. Collect the fractions you want Frac-950 and Frac-920 have the capacity to collect fractions in many kinds of vessels and over the whole flow rate range of ÄKTA design chromatography systems under the control of UNICORN. To achieve the best results it is important to preserve the resolution obtained from the column all the way to the collection vessel. The automatic peak fractionation functions in UNICORN enable the fraction collectors to work optimally. Fig 8. Fraction Collector Frac-950 (left) and (right) Frac-950 modified for collection of extremely small fraction volumes using tubing provided in Microfraction Collection Kit. 18-1153-57 AE 3

Technical specifications Frac-950 Operating data Max. flow rate 100 ml/min ph stability range 1 to 13, 1 to 14 (<1 day exposure) Fraction size Volume mode: 0.1 99999.99 ml Time mode: 0.1 99999.99 min No spillage range Dropsync. range: 0 2.0 ml/min Accumulator range: 2.0 100 ml/min Environment +4 C to +40 C 20% 95% relative humidity 84 106 kpa (840-1060 mbar) atmospheric pressure Frac-950 Tube racks Rack designation Tube Max number and color combination of fractions Rack A, yellow 18 mm tubes 120 30 mm tubes 8 Rack B, violet 12 mm tubes 240 Rack C, blue 96-well microplates 384 30 mm tubes 8 Rack D, red 30 mm tubes 45 Preparative mode Rack E, cream 30 mm tubes 80 Rack F, green 250 ml bottles 20 Funnel to Flask Funnels 30 (used with Rack E) 1 The following manufacturers microtitre plates are tested and approved by GE Healthcare to be used with this rack type: Greiner low: 655101, 651101, 650101. Greiner high: 780201. Nunc low: 143761, 168055, 156545, 163320 The differences between these articles are in the shapes of the bottoms of the wells and the inclusion of a lid in some cases. All articles have 96 wells. Frac-950 Physical data Degree of protection IP 21 Wetted materials PEEK (polyetheretherketone) UHMW-PE (ultra high molecular weight polyethylene) Glass Elgiloy HT Chemical resistance The wetted parts of the instrument are resistant to organic solvents and salt buffers commonly used in chromatography of biomolecules, except 100% ethyl acetate, 100% hexane and100% tetrahydrofuran (THF) Power requirement 100 120/220 240 VAC, 50-60 Hz (autorange switching) Power consumption 300 VA Dimensions, H W D 480 380 550 mm Weight 16.5 kg Frac-950 Ordering information Item Code no. Fraction collector Frac-950 (including Rack A) 18-6083-00 Fraction collector Frac-950 requires an ÄKTA design system controlled by UNICORN v 3.2 or higher. To use the preparative mode (requires UNICORN 4.0), order Fraction collector Frac-950 together with Prep Mode Conversion Kit 18-6083-18, and at least one of Rack E 18-6083-15 or Rack F 18-6083-16. Related product literature Data Files Code no. ÄKTAexplorer chromatography systems 18-1124-09 ÄKTApurifier chromatography systems 18-1119-48 UNICORN Control System 18-1111-20 ÄKTAmicro chromatography systems 28-9496-70 Accessories and consumables Item Quant./pack Code no. Microfraction Collection Kit 1 28-9487-80 Rack A, complete with bowl for 18 mm and 30 mm tubes 1 18-6083-11 Rack B, complete with bowl for 12 mm tubes 1 18-6083-12 Rack C, complete with bowl for 96-well microtitre plates and 30 mm tubes 1 18-6083-13 Rack D, complete with bowl for 30 mm tubes 1 18-6083-14 Prep Mode Conversion Kit 1 18-6083-18 (for use with Rack E and Rack F) Rack E, complete for 30 mm tubes 1 18-6083-15 Rack F, complete for 250 ml bottles 1 18-6083-16 Funnel to Flask Kit with funnels, tubing, and tubing organizer (for use with Rack E) 1 18-6083-17 PEEK tubing i.d. 0.25 mm, o.d. 1/16" 2 m 18-1121-36 PEEK tubing i.d. 0.50 mm, o.d. 1/16" 2 m 18-1113-68 PEEK tubing i.d. 0.75 mm, o.d. 1/16" 2 m 18-1112-53 PEEK tubing i.d. 1.00 mm, o.d. 1/16" 2 m 18-1115-83 Fingertight connector 1/16" 10 18-1112-55 Mains distribution cable, for connection to ÄKTA design 1 m 18-1032-08 UniNet cable 1.5 m 18-1117-75 UniNet cable 0.7 m 18-1109-74 UniNet cable 3 m 18-1109-75 UniNet cable 15 m 18-1117-74 EMC standards This device meets the requirements of the EMC Directive 89/336/EEC through the following harmonized standards: EN 61326 (emission and immunity) EN 55011, GR 2, Class A (emission) This device complies with part 15 of the FCC rules (emission). Operation is subject to the following two conditions: 1. This device may not cause harmful interference. 2. This device must accept any interference recevide, including interference that may cause undesired operation. 4 18-1153-57 AE

Technical specifications Frac-920 Operating data Max flow rate ph stability range Fraction size Environment Frac-920 Tube Racks 100 ml/min 1 to 13, 1 to 14 (<1 day exposure) 0.1 999.0 ml (0.1 ml increments) 0.1 999.0 min (0.1 min increments) +4 C to +40 C, 20% 95% relative humidity 84 106 kpa (840 1060 mbar) atmospheric pressure Rack Max. number Nominal tube Tube length of tubes diameter 12 mm 175 12 mm 50 180 mm 18 mm 95 18 mm 50 180 mm 30 mm 40 30 mm 50 180 mm Frac-920 Physical data Event mark 0.2 sec for both contact closure and TTL output in remote socket Protection class IP 22 Wetted materials PTFE (polytetrafluoroethylene) ECTFE (ethylenechlorotrifluoroethylene) Chemical resistance The wetted parts of the flow diversion valve, the bowl and the racks are resistant to organic solvents and salt buffers commonly used in chromatography of biomolecules, except 100% ethylacetate, 100% hexane and 100% tetrahydrofuran (THF) Power requirement 100 240 V AC, 50 60 Hz Power consumption 20 VA Dimensions 250 320 400 mm (H W D) Weight 4.5 kg Safety standard This meets the requirements of the Low Voltage Directive 73/23/EEC through the following harmonized standards: EN 61010-1 IEC 61010-1 CAN/CSA-C22.2 No. 61010-1 UL61010-1 EMC standards This device meets the requirements of theemc Directive 89/336/EEC through the following harmonized standards: EN 61326 (emission and immunity) EN 55011, GR 2, Class A (emission) This device complies with part 15 of the FCC rules (emission). Operation is subject to the following two conditions: 1. This device may not cause harmful interference. 2. This device must accept any interference recevide, including interference that may cause undesired operation. Frac-920 Ordering information Item Quantity Code no. per pack Fraction Collector Frac-920 complete with 18 mm tube rack 1 18-1177-40 Tube Racks, complete with bowl, tube support, holder and guide 12 mm 1 19-8684-03 18 mm 1 18-3050-03 30 mm 1 18-1124-67 Tube support 1 18-3054-02 Tube holder kit 1 18-8522-01 Tube spring, metal* 10 19-6057-01 Bowl 1 18-3051-03 Eppendorf tube holder for 12 mm rack 100 18-8522-01 Flow Diversion Valve, SV-923 1 11-0011-70 Tubing holder 1 18-6464-01 Funnel 1 19-6044-01 Funnel tubing 12/8, 250 mm 2 18-3020-01 Drive sleeve 5 19-6067-02 Mains distribution cable, 1 m 1 18-1032-08 Mains cable, US, 110 V~, 1 m 1 19-2447-01 Mains cable, EU, 220 V~, 1 m 1 19-2448-01 Remote control cable 1 19-6005-02 Event mark/ Signal cable 1 19-6006-01 Recorder/UV cable 1 11-0011-76 UniNet cable, 1.5 m 1 18-1117-75 Teflon tubing, i.d. 1/8, o.d. 3/16" 3 m 18-1112-47 Tubing connector for 3/16" o.d. tubing 10 18-1112-49 Ferrule for 3/16" tubing 10 18-1112-48 Stop plug, 5/16" 5 18-1112-50 Stop plug, 1/16" 5 18-1112-52 Union Luer female/1/16" male 2 18-1112-51 Union 1/16" female/m6 male 6 18-1112-57 Union M6 female/1/16" male 8 18-1112-58 PEEK tubing, i.d. 0.75 mm, o.d. 1/16" 2 m 18-1112-53 Teflon tubing, i.d. 0.75 mm, o.d. 1/16" 2 m 18-1119-74 PEEK tubing, i.d. 1.0 mm, o.d. 1/16" 2 m 18-1115-83 Fingertight connector 1/16" 10 18-1112-55 * Only for Tube rack 18 mm 18-1153-57 AE 5

For local office contact information, visit www.gelifesciences.com/contact www.gelifesciences.com/akta GE Healthcare Bio-Sciences AB Björkgatan 30 751 84 Uppsala Sweden imagination at work GE, imagination at work, and GE monogram are trademarks of General Electric Company. ÄKTA, ÄKTAexplorer, ÄKTAfplc, ÄKTAmicro, ÄKTApurifier and UNICORN are trademarks of GE Healthcare companies. All third party trademarks are the property of their respective owners. 2001-2009 General Electric Company All rights reserved. First published May 2001 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information. GE Healthcare UK Limited, Amersham Place, Little Chalfont, Buckinghamshire, HP7 9NA UK GE Healthcare Europe, GmbH Munzinger Strasse 5, D-79111 Freiburg Germany GE Healthcare Bio-Sciences Corp. 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327 USA GE Healthcare Bio-Sciences KK Sanken Bldg., 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073 Japan 18-1153-57 AE 06/2009