Science Together AZURA® Bio purification Extensive and flexible FPLC solutions 1 AZURA® Bio purification systems AZURA® Bio purification systems Complete solutions for FPLC on a minimum footprint: AZURA FPLC systems combine flexibility and Size Exclusion Affinity Ion-Exchange Hydrophobic Interaction reliability. The biocompatible/metal-free FPLC is the perfect choice for your protein purification task. Chromatography Chromatography Chromatography Chromatography (SEC) (AC) (IEX) (HIC) 1 3 Flexible & modular Intuitive software 2 PurityChrom® 4 Separate according Specific binding of Separation takes Separation is per- Easy scale-up Variety of detectors to size. protein of interest. place according formed based on See page 22 for a See page 23 for a to the charge of hydrophobic interac- specialized AZURA specialized AZURA the protein and tion and gradient system for SEC. system for AC. gradient elution. elution. Design your AZURA Bio purification system to A large range of different detectors make your your needs. Multiple functionalities such as auto-target molecules visible. Different flow rates and matic sample injection via autosampler, column compatibility to columns from all vendors offer switching, buffer and sample selection as well as maximum flexibility. The intuitive software fraction collection enable the user to automate PurityChrom® combines all the advantages of a the purification process. versatile purification software. Capture Intermediate Polishing Normally a combination of methods is used in protein purification. • The “capture” step purifies the protein from the crude extract. • The “intermediate” step removes further contamination. • The aim of the final “polishing” step is to get rid of all remaining impurities in order to gain a highly purified product. FPLC is a liquid chromatographic method for pu-sides, the method uses column materials out of • Automatization of two purification steps is possible using the especially rification of large biomolecules like proteins. Ex-agarose or polymer material which are very sen-designed AZURA Two step purification system (see page 24). ternal factors like high temperature, high pressure, sitive against pressure fluctuations and air bubbles. extreme pH, or solvents can disturb the protein We designed our systems to meet your puri-structure and are therefore avoided in FPLC. Be-fication challenges! 2 www.knauer.net/fplc 3 AZURA® Bio purification systems up to 8x up to 8x air sensor 5x bypass back A1 A2 pressure up to waste B1 B2 regulator 15x From simple to complex, from lab to pilot scale: Design your UV Con detector pH fraction collector AZURA® FPLC system according to your purification task! pump pump A B sample pump AZURA Bio Lab allows you to create FPLC systems with highest independence. Just pick your up to 8x modules and build-up the system yourself. The intuitive software PurityChrom will be flexibly adjusted to your system. Detection Various detectors: UV/VIS, diode array, refractive index, Intuitive fluorescence and a selection Sepapure® Automate your PurityChrom® of flow cells (see page 16) FPLC columns purification software (page 12) (page 20) (page 26) Fraction collection Various fractionation valves and fraction collectors with fraction racks ranging from 96-wel -plates up to bottles Injection (see page 18) Manual or automated injection: via loop, sample pump or auto- Suppporting sampler, sample selection valves all columns. available for up to 8 samples Buffer selection Cold-room Integrated buffer selection valve for 4 operation is buffers, extra buffer selection valve supported. optionally available (see page 6) Choose the Pilot series if you want to increase your productivity Configure your AZURA even more. Upscale our Lab configuration with same flexibility, soft-Bio purification system ware PurityChrom® and minimal footprint. Just transfer and upscale Find all FPLC products on Conductivity Column and loop Buffer delivery your methods. Flow rates up to 1000 ml / min. the following pages. monitor selection valve Quaternary or binary pumps Find more information: www.knauer.net With pH option Choose between five with flow rates up to (see page 16) columns or loops. 10 ml / min or 50 ml / min Reverse flow possible. (see page 6) 4 www.knauer.net/fplc 5 AZURA® Bio purification systems Pressure sensor with inline filter Buffer selection valve A1, A2, B1, B2 for binary gradient High-performance gradient pump P 6.1L HPG Precise and reliable pumps covering a wide flow rate range, optimized for low pulsation Ceramic pump heads: 10 or 50 ml/min gradient and buffer selection options. (400 or 200 bar) Low-volume mixer With or without degasser Automated switching between buffers is important for method development, column cleaning and regeneration. The pump P 6.1L features a build-in 2 x 2 buffer selection valve (A1, A2 and B1, B2) or P 6.1L LPG 4 x buffer selection valve (A, B, C, D). Buffer blending Let the You can extend buffer selection with additional valves each for up to 8 buffers. valve A, B, C, D pump mix for quarternary gradient your buffer. Ceramic pump head: 10 ml/min (400 bar) 8x 8x 8x 8x 8x 8x 8x 8x D D A A A1 A1 A2 A2 8x 8x 8x 8x 8x 8x 8x 8x Pumps for high flow rates C C B B B1 B1 B2 B2 400 pump pump pump pump pump pump 200 ar) A AB B (b Titanium pump head 100 AZURA pump P 6.1L LPG - Quaternary gradient AZURA pump P 6.1L HPG - Binary gradient ssure with optional gradient valve Pre 50 100 250 500 1 000 Pressure sensor Flow rate (ml/min) Gradient options Ceramic pump head From binary to quaternary gradient, with 10 or 50 ml/min additional P 2.1L pumps or cost-effective Isocratic pump with small footprint for binary low pressure gradient (2 × 1 buffers, dedicated applications or sample loading. A B A B C Pressure sensor up to 800 ml / min) or ternary low pressure gradient (3 × 1 buffers, up to 220 ml / min). pump pump Binary low-pressure gradient Ternary low-pressure gradient 6 www.knauer.net/fplc 7 AZURA® Bio purification systems A flexible combination module A quaternary low pressure gradient (LPG) module* ferent channels. The buffer in a binary high pres-dynamically composes the buffer on the inlet-side sure gradient (HPG) system is composed by The Assistant ASM 2.2L is a docking station for Depending on the integrated modules the assis-or low pressure side of the pump head, by quick-combining the flow of two pumps. three compact devices. Valves, pumps and UV tant fulfills many different tasks like eluent delivery, ly switching the selection valve between the dif-detectors can be combined in one housing. The detection, sample and solvent selection, sample plug-in modules are removed by loosening four injection, column switching or fraction collection. screws allowing the user to exchange modules in An assistant including a pump, injection valve, case of service within minutes. Likewise, the con-and detector features a complete FPLC system, Quaternary Binary figuration of the FPLC system can be adapted to like AZURA Bio SEC or AZURA Bio AC. As a part gradient gradient new requirements. Routine maintenance work of a larger system, the ASM 2.2L allows the user e.g. replacing the lamp of a detector is easily per-to customize the system configuration according • Low investment costs • Less wear formed by the user. to the purification challenge. • Limited flow rate range • No flow rate limitation • Channel usable • Sample pump for sample for sample injection injection (recommended) Freely combine pumps, valves • Gradient accuracy abso- • High accuracy for special and detectors in one housing lutely sufficient for FPLC application mAU 250 200 24% 25% 23% 22% 21% 150 20% 100 Use the web-based assistant configurator to find your 50 Time desired AZURA® ASM 2.2L module combination: Excellent gradient reproducibility of 0.3 % RSD. overlay of 6 repetitions at 1 ml / min run with pump P 6.1L www.knauer.net/assistantconfigurator low pressure gradient version *For higher flow rates available as ternary or binary gradient (see page 7). 8 www.knauer.net/fplc 9 AZURA® Bio purification systems Choose between manual or automated sample injection. Different options for column selection are available. Available modules include injection valve, sample pump, loop valves and autosampler. ass 2x • Select two columns or one column and one bypass pby • Flow rates up to 500 ml / min possible Integrated into assistant or as standalone module: The Multi-Injection valve for AZURA Bio Lab systems enables the sample injection via loop and sample pump. Using this smart valve the setup of an two-step purification is fast and easy. • For up to 5 columns and 1 bypass s ow as from pump to column • Reverse flow yp 5x The 2-position injection valve for AZURA Bio pilot systems is b erse fl • Flow rates up to 50 ml / min perfect for injection via a sample loop or a sample pump. If both rev injection possibilities are needed simply add two valves to your configuration. Why is the reversed flow option popular in 1,600 Integrated into assistant or as standalone module: to affinity chromatography? The AZURA P 4.1S is perfect for injecting large sample volumes and injection valve In affinity chromatography your target molecules repetitive loop fills. will accumulate at the top of the column. Elution 1,200 Higher concentration in the same direction dilutes your target molecule )UA Do you have many samples? along the column. By elution with reversed flow (m 0 8 800 up to 8 x You can extend your configuration with additional valves for 8 samples. you increase the concentration while decreasing A 2 the sample volume. 400 5x The option has two major advantages. Clean your The Loop valve can be optionally used to connect up to 5 loops to columns more efficiently using reverse flow. By the system or to collect intermediate fractions in automated two-this you elute contamination the shortest way and 0 step purifications with an AZURA Bio Lab system. Various sample from minimize damage to the column. 70 75 80 loops are available. injection valve to Volumen (ml) injection valve Less elution volume Process many different samples fully automatically with the Autosampler AS 6.1L. Use the column selection assistant to select eight ow • Up to 10 ml injection volume columns assuring a flow rate up to 500 ml/min. 8x • From microtiter plates to standard vials An additional valve allows to reverse the flow. erse fl • Active cooling rev • Fully supported by PurityChrom® software • Metal-free 10 www.knauer.net/fplc 11 AZURA® Bio purification systems Developed for your FPLC application Reliable and convenient The perfect addition to any FPLC system In size exclusion chromatography biomolecules are seperated according to their size. There are two In affinity chromatography a higly specific interac-off in the wash phase. The elution of the target different methods used in SEC which are defined by the matrix of the FPLC columns. tion between the biomolecule of interest and the biomolecule is realized by washing the column with column matrix is resulting in the enrichment of the a buffer including a high amount of competing biomolecule at the stationary phase during the ligand or low pH. Group separation High resolution separation loading phase. Byproducts can be easily washed Separation of small molecules Separation of larger biomolecules from large molecules within the fractionation range of the Recombinant tagged proteins Antibodies and antibody fragments (e.g. Desalting) column matrix His – Tag via Ni-NTA column via Protein A immobilized on column GST – Tag via Glutathione column via Protein G immobilized on column Large Small BSA molecules molecules U) NaCI U) A A m m U) al ( al ( A n n m Sig Sig al (n Sample loading/binding of biomolecule Sig Wash step Elution of biomolecule Volume (ml) Volume (ml) Re-equilibration of column Volume (ml) Prepacked 1 ml or 5 ml columns Prepacked 1 ml or 5 ml columns Key features Key features • Dextran based beads with particle sizes ranging from 20 – 50 µm • Agarose based beads with particle size of 100 µm on average • 5 kDa exclusion limit (all molecules bigger than 5 kDa are not retained) • Static binding capacity: Ni-NTA > 40 mg/ml; Glutathione > 10 mg/ml; • Recommendend flow rates: 0.5 – 2 ml/min (1 ml column); 1 – 5 ml/min (5 ml column) Protein A > 30 mg/ml human IgG; Protein G > 15 mg/ml human IgG • Maximum pressure: 3 bar • Recommendend flow rates: 0.5 – 2 ml/min (1 ml column); 1 – 5 ml/min (5 ml column) • Stored in 20% Ethanol • Maximum pressure: 3 bar • Stored in 20% Ethanol 12 www.knauer.net/fplc 13 AZURA® Bio purification systems In ion-exchange chromatography biomolecules biomolecules are binding to a negative column are separated according to their charge. Anion matrix. The bound molecules are released from exchange is the method in which negatively the matrix by a gradual increase in ionic strength charged molecules are binding to a positive ma-of the elution buffer. trix and in cation exchange positively charged Comparison to other vendor See information on detailed Anion Exchange Cation Exchange comparison of columns: Strong Anion Exchanger (Q) Strong Cation Exchanger (SP) www.knauer.net/sepapure Q SP N+ SO -3 Weak Anion Exchanger (DEAE) Weak Cation Exchanger (CM) - Next to prepacked FPLC columns KNAUER also cation. In addition to the media used with the OH + O O O NH CM offers FPLC bulk media for high performance pu-prepacked FPLC cartridges we also offer SEC DEAE O rifications from lab to large-scale protein purifi-resins for high resolution separations. u] Resin Type / 5 ml 10 ml 25 ml 50 ml 100 ml 150 ml 250 ml 500 ml 1000 ml Volume gnal [mASi u] Glutathione • • • • • • u] S] gnal [mA U) Si Ni-NTA • • • • • A gnal [mA m Si vity [mcti al ( u n Protein A • • • • • • nd Sig oC Protein G • • IEX-Resins • • • • Volume (ml) SEC 75 • • • • • • • SEC 200 • • • • • • • Prepacked 1 ml or 5 ml columns Key features Key features • Agarose based beads with particle size of 100 µm on average • Cross-linked agarose-dextran composite with a particle size of 35 µm on average • Ion capacity: 0.08 – 0.16 mmol/ml • Maximum pressure: 3 bar (SEC 75) or 4 bar (SEC 200) • Recommendend flow rates: 0.5 – 2 ml/min (1 ml column); 1 – 5 ml/min (5 ml column) • Separation range of Sepapure SEC 75: 3 – 70 kDa • Maximum pressure: 3 bar • Separation range of Sepapure SEC 200: 6 – 600 kDa • Delivered in 20% Ethanol • pH tolerance: 2 – 14 (short term) / 3 – 12 (long term) 14 www.knauer.net/fplc 15 AZURA® Bio purification systems Refractive Index Detector for cost-effective, fast Universal and reliable analysis of non-UV absorbent com-detection We provide a choice of UV/VIS pounds. detectors, ranging from single variable wavelength to 8-channel diode array detector with 3D scan capability. A wide range of third-party detectors can be seamlessly integrated into AZURA® systems. Light scattering detector Detector UVD 2.1S MWD 2.1L DAD 2.1L Gradient Using the unique Low Temperature technology, Compact and cost-ef-Reliable multichannel Diode array detector compatible this Evaporative Light Scattering detector LC al-fective variable single UV/VIS detector for peak purity check and universal lows universal high sensitivity detection of non-UV wavelength UV/VIS detection active substances. detector Wavelength 190 – 500 nm 190 – 700 nm 190 – 700 nm Caution! May cause Channels 1 4 8 higher back pressure Fluorescence detector RF-20A 3D scan n/a n/a + The fluorescence detector RF-20A provides Sensitive world-class sensitivity, excellent maintainability and selective Integrable in ASM + and diverse validation / support functions. It sup-fluorophor detection ports a wide range of applications from conven-More UV detectors available for your applications: www.knauer.net/detectors tional to high-performance analysis. AZURA® Conductivity Monitor CM 2.1S • Conductivity monitor for checking salt gradient The KNAUER interface box IFU 2.1 LAN allows • Flow rates up to 100 ml / min precise analog data acquisition of third party • 0.01 mS/cm-999 mS/cm modules. • pH option available Flow cells for CM 2.1S Caution! May cause Analytical 1/16" 10 ml / min 160 bar 30 µl volume higher back pressure Preparative 1/16" 100 ml / min 100 bar 300 µl volume Select from an impressive range of easily tially from the device providing enhanced secu-exchangeable flow cells which cover a wide range rity for hazardous, explosive or toxic work pro-of application. Optional fiber optics technology cesses. offers the possibility to separate the flow cell spa-16 www.knauer.net/fplc 17 AZURA® Bio purification systems Collect large quantities or large numbers of fractions Accessory Features Benefit Manually – collection by direct control Volume-based – collection at defined volumes Pressure Control • Contains two pressure sensors Monitors pressure over the column bed and protects Peak-based – collection according to detector signal • Automatic determination of column from damage pressure difference with Purity- Chrom® Outlet Valves 16x • Connect 1/16” or 1/8” tubings • Collecting large quantities ideal for flow- • Up to 250 ml / min and 60 bar throughs and wash fractions Air Sensor • Detect end of buffer or end of Protect column from air • Used as single unit for up to 16 fractions in an sample with PurityChrom® damage and support AZURA Bio Lab system or 8 fractions in an automation ( e. g. sample • Up to four air sensors per system AZURA Bio Pilot system injection) 15x waste • For transparent tubings with • Valves can be daisy chained with a fraction col-1/16” or 1/8” or 1/4” outer lector diameter AZURA® Click • Attach air sensor, pressure Organize your system. fraction collector control, AZURA Organizer or your interface box to the side panel of your AZURA L device AZURA® Organizer • Attach columns from 5 mm to Organize accessories 26 mm diameter, falcon tubes, a directly at the system and back pressure regulator or a pH reduce dead volume flow cell Foxy Fraction collector Vario 4000 & Vario 4000 plus Back pressure • Apply a constant back pressure Prevent formation of air The Foxy R1 and Foxy R2 are versatile fraction The Vario 4000 is a more advanced fraction regulator (BPR) to your system bubbles after the column collectors which fit to every purification need. collec tor for demanding applications with high • Freely adjustable between 1-20 which disturb detector signal flow rates and a high number of fractions. Indivi-bar or • Up to 125 ml / min for Foxy R1 and 1 000 ml/min dual rack types are programmable. Just assemble 20-103 bar for Foxy R2 your rack to your needs. • Wide choice of racks from 96-well microplates AZURA® • Install AZURA systems at space-lim- Space-saving solultion for up to bottles or funnels • For flow rates up to 1 000 ml / min Benchtop Rack ited sites, especial y in cold rooms. AZURA system setup • Double capacity for Foxy R2 with automatic • High number of fractions rack recognition • Supported in software Puritychrom® • Active cooling for Foxy R1 • Standalone operation possible • Supported in software Puritychrom® • Stand-alone operation • Repeated collection in same vials 18 www.knauer.net/fplc 19 AZURA® Bio purification systems Components from lab to pilot Product Features Page Buffer delivery Compact pump 10 or 50 ml / min, isocratic 6 Product Features Page Gradient pump 10 or 50 ml / min, quaternary: selection of 4 buffers (A, B, C, D) 7 Binary: selection of 2 buffers (A1, A2, B1, B2) AZURA Bio SEC 0.001–10 ml/min, maximum 150 bar, injection valve sample for 22 sample loops, variable single wavelength UV-detector, XY Scale-up pump 100, 250, 500, 1000 ml / min, binary to quaternary gradient 7 fraction collector, PurityChrom® software Extended buffer With additional valves each for 8 buffers 6 selection AZURA Bio AC 0.01–50 ml/min, maximum 150 bar, selection valve for 8 buffers/ 23 Sample selection For maximum 8 samples 10 samples, variable single wavelength UV-detector, fraction valve for 7 fractions and waste, PurityChrom® software Columns Column selection For 2 columns, 5 columns and 5 columns with reverse flow 11 AZURA Bio Lab 0.01–50 ml/min, maximum 50 bar, sample injections via sample 4 valves option loop and sample pump, variable single wavelength UV-detec-Sepapure® columns Columns and media for Affinity, Size Exclusion and Ion-tor, XY fraction collector, PurityChrom software (basic license). 12 and media Exchange Chromatography Exemplary System - confiugre your FPLC system based on your purification requirements Detection Wide choice of Variable single wavelength UV, multiple wavelength UV, 16 AZURA Bio Lab 0.01–50 ml/min, maximum 50 bar, sample injection via sample 24 detectors full spectra diode array (DAD/3D Spectrum), conductivity and Advanced configuration loop or sample pump, automated storage and reinjection of pH monitor, fluorescence, refrective Index proteins possible , multichannel UV-detector, conductivity Fraction collection detector, outlet valve , XY fraction collector, PurityChrom Software Fractionation valve For 6 to 16 fractions, depending on the valve type with flow-18 rates up to 1000 ml/min Fraction collector From 96-well microplates up to bottles or funnels, 18 AZURA Bio Pilot Up to 1000 ml/min, sample pump for large sample volumes, 5 up to 1000 ml / min variable single wavelength UV-detector, XY fraction collector, Sample injection PurityChrom® software in basic configuration. Configure your FPLC system based on your purification requirements. Scale-Injection valve 1/16” tubing: up to 50 ml / min 1/8” tubing: up to 500 ml / min 10 up is possible with same flexibility, software but minimal Sample pump 10 or 50 ml / min 10 footprint. Autosampler Up to 10 ml injection volume, from microtiter plates to 10 ml vials 10 Software PurityChrom® Highly flexible method writing, intui tive user-interface, volume-26 software or time-based, with special features like system visualisation, hold & adjust option, extended threshold functions, check for impurities Safety features Accessories for protec- Air sensor, pressure control, back-pressure regulator, leak 19 tion and automation manage ment, mounting solutions 20 AZURA® Bio purification systems Time consuming gel filtration runs? For affinity chromatography AZURA Compact SEC systems take over time-con-ease of use. Pre-designed methods are included The AZURA Compact AC system qualifies for fast the selection valve. Your proteins of interest are suming SEC methods in your lab without blocking in the software for the designated SEC system and reliable affinity chromatography. Select your detected by UV and automatically collected via your valuable FPLC system. Due to its compact and can be easily adapted by changing the column sample, your washing and elution buffer using the fractionation valve. design and intuitive FPLC software PurityChrom®, volume. AZURA Compact SEC supports al columns the system offers outstanding performance and available on the market. waste 8x 8x UV waste pump column detector UV pump injection column detector fraction collector Use the selection valve for your buffers and sample. Key features • Flow rate: 0.001 – 10 ml / min; 0.1 – 8.0 ml / min (recommended) Key features • Maximum system pressure: 150 bar • Automatic sample/ buffer selection valve for • Injection valve for sample injection 2,000 Binding Washing Elution up to 8 buffers or samples via sample loop • Fraction valve (8 ports) for fractionation • Variable single wavelength • Flow rate: 0.01 – 50 ml / min; 1 – 40 ml / min 1,600 UV-detector (190 – 500 nm) (recommended) • Fraction collector for fractionation • Variable single wavelength 1,200 • Columns from all vendors can be used ) UV-detector (190 – 500 nm) U • PurityChrom® software A • Columns from all vendors can be used 800 (m 0 8 • PurityChrom® software A 2 • Maximum system pressure: 150 bar 400 0 Protein purification based on high affinity 5 10 15 20 25 30 Chromatogram & Legend Volume (ml) 22 www.knauer.net/fplc 23 AZURA® Bio purification systems up to 8x up to 8x air sensor 5x bypass back A1 A2 pressure up to waste B1 B2 regulator 15x Special multicolumn chromatography solutions UV Con detector pH fraction collector pump pump A B AUTOMATION Protein purification involves most of the times two to three steps: sample pump 1. capture step up to 8x 2. optional intermediated step 3. polishing step The transition from one to another step generally chromatographic purification steps into one meth-involves manual interaction and thus is time con-od eliminates manual sample handling and min-suming. Automation by combining these steps imizes time spent between steps. This automation increases the efficiency and optimizes the work-strategy can be easily adapted to each purification Automated two-step purification of mouse IgG antibodies flow. The quick and automated linkage of multiple task. 2,000 Phase 1 Phase 2 Phase 3 Phase 4 A Detection Multiple wavelength detector Sample injection For large and small 1,500 sample volumes )UA Conductivity (m 0 8 Monitor your salt gradient A 2 1,000 B1 Column selection Outlet valve Select between 5 columns Collection of large 500 and one bypass, volume fractions reverse flow B2 like flow through, wash fractions etc. 0 Fraction Buffer selection 0 10 20 30 40 50 Collection Select between Volume (ml) 4 buffers The affinity chromatography step was automatically combined with a gel filtration step to exchange the buffer of the purified mouse IgG antibodies; Phase 1: Column equilibration, Phase 2: Sample injection and washing, Buffer delivery Phase 3: Elution of IgG from protein A column, Phase 4: Desalting of IgG System pump with binary gradient 24 www.knauer.net/fplc 25 AZURA® Bio purification systems Extended threshold functions Automate any software function triggered by signals of any channel. Create methods with highest flexibility to realize Automatically start fraction collection at the complex applications. Offline licenses for creating beginning of your desired peak. Protect the sys-methods and evaluating data are free of charge. tem from overpressure and air bubbles. After end of sample detection the software offers the possibility to automatically start or continue the run. Automate the whole purification starting from sample injection, via column washing to elution. Check for impurities – System visualization full spectra diode array (DAD) Keep an eye on your system by using the system visualization. The interactive flow path allows to Check the purity of your peaks based on the ab-control your system. Switch valves, start pumps, set autozero, start fraction collection and more with sorbance spectra anywhere in the elution profile. just one click. Hold & adjust (a running method) Stay always in control and change the parameters on the fly, even during a progressing method. Get familiarized with manual y controlling your system, writing methods and analyzing your data using PurityChrom®. www.youtube.de/KNAUERhplc 26 www.knauer.net/fplc 27 AZURA® Bio purification systems Intuitive data analysis Integrate peaks fully automatically or manually. Receive the peak results by clicking on one button. AZURA® Bio purification solution by KNAUER „Our KNAUER FPLCs are the workhorses in the lab. ” “My lab studies the structure and function of membrane proteins. Due to the inherent instability of these proteins we pu-rify them in the cold room. We needed robust FPLCs with good pumps that tolerated these conditions well. In addition, the systems needed to be easy to maintain. Knau-Solvent supply – calculate the consumption of buffers er provided us with skilled advice on virtually every component The solvent supply function calculates the con-A B C D Waste of the system, ranging from tubing and pumps up to the soft-Jun. Prof. Dr. Eric R. Geertsma sumption of buffers and the waste level for the ware. Consequently, our systems are perfectly tailored to our Institute of Biochemistry, current run, thus preventing the column from needs. Most of the maintenance we can do ourselves. For Goethe-University Frankfurt running dry and flooding the lab. remaining questions, we can rely on the great support Knau-Photo: Uwe Dettmar er offers. Our Knauer FPLCs are the nononsense workhorses in the lab. I highly recommend Knauer.“ Stacked Injection Size exclusion chromatography separates the proteins according to their size. After selection System components of SEC medium, sample volume and column di- • AZURA® UV Detector UVD 2.1S mensions are the two most critical parameters • AZURA® Valve Drive V 2.1S that will affect the resolution of the separation. 25% • AZURA® Pump P 4.1S For most SEC runs the sample volume should not time saving • Foxy fraction collector exceed 2% of the total column volume to achieve maximum resolution. U)Am al (n For larger sample volumes the sample must there-Sig AZURA Compact SEC systems fore be divided into different runs. However, this take over time-consuming SEC takes a lot of time and is not very efficient. With methods in your lab without the stacked injection function in PurityChrom it blocking your valuable FPLC is possible to run different runs automatically one system. after the other. The injection of the next run takes place during the current run, so that the time un-Contact us: til the elution of the first peak can be fully exploit-sales@knauer.net ed. This increases efficiency and saves time. Volume (ml) 28 www.knauer.net/fplc 29 M E THOD SEC AC IEX HIC Size Exclusion Affinity Ion-Exchange Hydrophobic Interaction Chromatography Chromatography Chromatography Chromatography up to 8x up to 8x up to 8x up to 8x air sensor 5x Worldwide partner in air sensor bypass 5x back A1 A2 science since 1962 pressure up to waste B1 B2 bypass back A1 A2 pressure regulator up to 15x waste B1 B2 regulator 15x UV Con UV Con detector pH fraction collector detector pH fraction collector pump pump Based in Berlin, KNAUER is a medium-sized, ow-tems and components for analytical HPLC / UHPLC, pump A pump B sample ner-managed company that has been serving the preparative HPLC, fast protein liquid chromato-A B sample pump pump sciences since 1962. We develop and manufac-graphy (FPLC), multi-column chromatography / up to 8x up to 8x ture scientific instruments of superior quality for simulated moving bed (SMB), and osmometry. liquid chromatography. The range includes sys-BUFFER SELECTION SAMPLE COLUMN SELECTION DETECTION FRACTION & DELIVERY INJECTION COLLECTION 10 ml/min binary Multi-Injection valve Column selection UV/VIS Outlet valve gradient pump P 6.1L valve up to 50 ml/min single wavelength ..... x Injection valve (5 columns, one Foxy fraction 10 ml/min quaternary bypass, reverse flow) UV/VIS multiwave- collector with pump P 6.1L Sample pump length fixed rack types Independent and .......module Column selection family owned 50 ml/min binary (two columns or Conductivity Labocol fraction gradient pump P 6.1L Sample selection one bypass) collector with ......valve: ..... x inlets pH individual rack types ..... x 100 ml/min pump P 2.1L Column selection Biocompatible high flow Fluorescence Rack for fraction ..... x 250 ml/min pump P 2.1L ......Autosampler AS 6.1L (5 columns, one collector The founder Dr. Herbert Knauer and his wife owner of the company since 2000. Several bypass) Refractive index ..... x 500 ml/min pump P 2.1L Roswitha are still active as advisers in the com-awards for outstanding products and innova-Column selection Light Scattering pany to this day. The couple‘s daughter, Alexand-tions as well as entrepreneurial excellence make ..... x 1 000 ml/min pump P 2.1L high flow (5 columns, one Analog integration ra Knauer, has been managing director and KNAUER a „leading employer“. Ternary gradient module bypass, reverse flow) of further detectors ......for pump P 2.1L Binary gradient module ......for pump P 2.1L COLU M N S & M E DI A ..... x Buffer selection valve SEC: Desalting ..... ml AC: Protein A ..... ml IEX: DEAE – Weak anion exchange ..... ml We separate molecules ......(8 further inlets) SEC: SEC 75 ..... ml AC: Protein G ..... ml IEX: CM – Weak cation exchange ..... ml SEC: SEC 200 ..... ml AC: Ni-NTA ..... ml IEX: Q – Strong anion exchange ..... ml and unite people. AC: Glutathione ..... ml IEX: SP – Strong cation exchange ..... ml ACCESSORIES www.knauer.net ..... x Air sensor ..... x Air sensor Pressure control ..... x Back pressure AZURA Organizer main pump feed pump (2 pressure sensors) regulator ..... x Tubing 1/16” ..... x Tubing 1/8” ..... x Tubing 1/4” Workstation (Windows) 30 www.knauer.net/fplc Technical data or prices are subject to change without All trademarks are the property of their respective owners. notice. Prices may vary by country and do not include Our general terms and conditions apply: taxes, customs duties or delivery. www.knauer.net/terms Distributed by: 20 S/0.5/02. 7855U/V20 ER 20UAN K© KNAUER +49 30 809727-0 Wissenschaftliche Geräte GmbH +49 30 8015010 (Fax) Hegauer Weg 38 info@knauer.net 14163 Berlin, Germany www.knauer.net