MediaScout Customized to the specific needs of our clients

Transcription

1 MediaScout Catalogue 2008 mediascout

2 mediascout MediaScout Customized to the specific needs of our clients MediaScout Is the design technology that makes these products suited to the specific needs of our clients, providing a choice of a predefined or a customized set of commercially available process separation resins prepacked into ready-to-use columns of various formats. Atoll products enable customers to obtain valid results in shorter timeframes which confers a valuable competitive advantage Atoll offers outsourced column packing, so users can focus attention on their application workload All the MediaScout column products can be packed to high efficiency with any resin type (>10 µ particle size) MediaScout MiniColumn parallel chromatography High Throughput sample handling A high throughput screening tool in a mini column array format, comprising a base plate designed in the standard 96-well microplate format and a specified number of up to 96 individually prepacked MiniColumns, filled with any commercially available bioprocessing resins. Generate your customized MiniColumn array at our online shop. MediaScout MiniChrom Miniaturized low to medium pressure chromatography columns, for use in bioseparation applications, with small sample volumes in various geometries. All column parts are made from mechanically and chemically stable polymers, predominantly from polypropylene. Generate your customized MiniChrom column library at our online shop. MediaScout MaxiChrom MaxiChrom, disposable and totally incinerable cgmp produced columns for special applications. For example packed with a suitable resin as disposable item for removal of detergents after virus inactivation during production in completely incinerable columns. Or packed as disposable guard column to protect large valuable process units. MediaScout ValiChrom Purpose designed prepacked columns packed with your own specified resin material for scaledown validation studies. Professionally cgmp-packed into precision bore glass columns with accurate compression to represent conditions in your production column for a price that cannot be matched. Provide a valid small scale model of large process columns to replicate the process scale unit s performance criteria. Atoll will pack any resin of your choice or your own resin into columns with the length to match the purification scheme of the customer, preferably 1cm 2 cross section. Lot-to-lot validation studies Virus or endotoxin clearance CIP validation studies Multi cycle resin lifetime studies MediaScout ResiQuot A vacuum manifold device for the accurate and reproducible preparation of aliquots from solids of suspensions into any kind of 96-well micro plates. Subsequently, these are used in experimental determination of chromatographic parameters in batch mode for example batch kinetics, batch isothermes and other applications. Avoids inaccurate metering created by other particulate sampling methods such as pipette, settlement etc. Table of content MiniColumn 4 Why 3 different design are offered 4 Starter Kit 5 Pre-set Kits 5 Individually customized Starter Kits 6 PipetColumn 7 CentriColumn 7 RoboColumn 8 Technical Information 9 Ordering Information 9 MiniChrom 10 Technical Information 13 Ordering Information 13 ValiChrom 14 Technical Information 15 Ordering Information 15 MaxiChrom 16 Technical Information 17 Ordering Information 17 ResiQuot 18 Technical Information 20 Ordering Information 20 Frequently asked questions 21 Applications 22 Price List 23 To view up-to-date prices, please see (SUPPORT-DOWNLOADS) 2

3 mediascout mediascout MiniColumn MiniChrom Operation mode: ready-to-use LC columns MaxiChrom Operation mode: ready-to-use LC columns CentriColumn Operation mode: 96 well format ID L Volume ID L Volume ID L Volume mm mm ml mm mm ml mm mm ml ,2 11,3 x x 5 2,5 0, ,5 32 x x 5 5 0, x x ,2 100 x x , PipetColumn Operation mode: 96 well format ID L Volume mm mm ml 5 2,5 0, , ,2 RoboColumn Operation mode: 96 well format ID L Volume mm mm ml 5 2,5 0, , , ,6 StarterKit Operation mode 96 well format ID L Volume mm mm ml ,2 ValiChrom Operation mode: ready-to-use LC columns ResiQuot Operation mode: metering of slurried particles ID L Volume mm mm ml 11,3 x x 11, ,

4 mediascout MiniColumn MediaScout MiniColumn for rapid resin screening and methods development This high throughput (real parallel chromatography) screening tool is an array of individual small columns installed on a base plate conforming to the standard 96 well microplate format. Features Identical column geometry Using any resin Compression packed Parallel chromatography Benefits Evaluation of base parameters, valid comparable results Broad range testing scope Optimized efficiency/ performance Very high productivity, rapid resin and method screening MediaScout is the design technology that enables our products to be customized to the specific needs of our clients, providing a choice of a predefined or a selected set of commercially available process separation resins prepacked into ready-to-use MiniColumns. Standardized chromatographic tests provide relative performance data of different resins so they may be checked in parallel in the shortest time frame possible. Columns are prepacked with commercially available separation media according to the needs of the user. Units may be sealed with a flexible capping mat at both ends for storage and reuse. Material from MiniColumn arrays can be collected in any standard 96 array plate. MiniColumns may be operated either by positive liquid displacement pipette, centrifuge or by robotic fluid handling equipment. 3 different MiniColumn versions available, dedicated for liquid flow by Centrifuge R CentriColumn Positive displacement pipette R PipetColumn Robotic fluid handling equipment R RoboColumn Operation: Manual or robotic pipetting or centrifugation to move liquid MiniColumn: 1 CentriColumn 2 PipetColumn 3 RoboColumn Small packed columns with separation media in a 96-array format. Sizes from µl available. Sample reservoir: 200µl 100µl 50µl resin bed 230µl 280µl 315µl sample chamber 4

5 media MiniColumn MediaScout MiniColumn Why do Atoll provide 3 different versions? 1. The CentriColumn has a reservoir insert above of the resin bed to hold analytes before liquid is moved through the resin. The outlet is a male luer conus.the centrifuge version assures a synchronous passage of liquid through up to 96 columns into a standard 96 or deep well plate. For applications when a large number of samples must be processed but when a fixed tip liquid handling station is not available. 2. The PipetColumn version has an inlet piece that fits most tips of positive displacement pipettes (Brand, Eppendorf, Gilson, etc.). From the inlet a capillary leads to the resin and a capillary outlet ensures a liquid flow as in a LC system. Liquid flow is controlled by the pipette, either manually or by a motorized version into a 96 deep well plate. For applications that need flow control and when a fixed tip liquid handling station is not available. For checking out the MiniColumn system Atoll offers a Starter Kit. 3. The RoboColumn version has an inlet that seals to fixed tips of TECAN s (and some others*) robotic liquid handling stations. From the inlet a capillary leads to the resin top and a capillary outlet on the bottom ensures a liquid flow as in a HPLC system. Flow is controlled by the TECAN station into a 96 deep well plate. For fully automated applications with high sample throughput and accurate control of flow rate and transfer volumes. MediaScout Starter Kit First steps in resin screening/method development In order to evaluate and try out the system, Atoll offers a Starter Kit which enables you to develop a screening protocol on an 8-row module, which can be then transferred to full 96 array operation. This Starter Kit can be seen as the simplest chromatographic system in the world. The Starter Kit consists of a row of 8 columns containing different media (listing below) stacked into a holder. This is positioned on a frame on top of a standard UV-readable 96-wellplate (e. g. UV-Star, Greiner). How to operate the Kit? Using a positive liquid displacement pipette: Set pipette into dispense mode. Regulate the speed to about 0.5 ml per minute Brand Handy Step Eppendorf Xstream Gilson Repetman 1. Prepare the appropriate buffers. 2. Condition all columns with starting buffer 3. Apply sample(s) by pipette 4. Elute with appropriate buffer into adjacent wells by shifting the 8-row along the 12 positions 5. Read results in a plate reader By this method 12 binding or respective elution conditions can be checked. To optimise buffer/solvent usage and preparation, each chromatographic mode may require a corresponding Kit. * Check with Atoll to confirm compatibility 5

6 media MiniColumn Starter Kits Customized Kits using your own choice of resins (Please contact Off-the-shelf Kits (see below) MediaScout Starter Kits Q-Kit 1 GE-Healthcare Capto Q 2 GE-Healthcare Q-Sepharose FF 3 BioRad UNOsphere Q 4 Merck Fractogel EMD TMAE Hicap M 5 Merck Fractoprep TMAE 6 Tosoh Toyopearl Super Q (M) 7 Pall-BioSepra Q Ceramic HyperD F 8 Applied Biosystems Poros 50 HQ S-Kit 1 GE-Healthcare Capto S 2 GE-Healthcare SP-Sepharose FF 3 BioRad UNOsphere S 4 Merck Fractogel EMD SO3 M 5 Merck Fractoprep SO3 6 Tosoh Toyopearl MegaCap II SP 550 C 7 Pall-BioSepra S Ceramic HyperD F 8 Applied Biosystems Poros 50 HS HIC-Kit 1 GE-Healthcare Phenyl Sepharose 6 FF high sub 2 GE-Healthcare Phenyl Sepharose 6 FF low sub 3 GE-Healthcare Butyl-S Sepharose 6 FF 4 Tosoh Toyopearl Butyl 650 M 5 Tosoh Toyopearl Super Butyl 550 C 6 Tosoh Toyopearl Phenyl 650 M 7 Merck Fractogel EMD Propyl S 8 BioRad MacroPrep t-butyl Protein A-Kit 1 GE-Healthcare MabSelect 2 GE-Healthcare MabSelect SuRe 3 GE-Healthcare MabSelect Xtra 4 GE-Healthcare Protein A Sepharose CL-4B 5 Millipore PROSEP -va HC 6 Millipore PROSEP -va Ultra 7 Pall-BioSepra Protein A Ceramic HyperD F 8 Applied Biosystems Poros 50 A IMAC-Kit 1 GE-Healthcare Chelating Sepharose FF 2 GE-Healthcare IMAC Sepharose FF 3 Merck Fractogel EMD Chelate M 4 BioRad Profinity IMAC 5 Pall-BioSepra IMAC HyperCel 6 Tosoh Toyopearl AF-Chelate-650M 7 Sterogene Nickel Chelating Superflow 8 Sterogene Cobalt Chelating Superflow HA-Kit 1 BioRad Macro-Prep ceramic hydroxyapatite Type I 40 µm 2 BioRad Macro-Prep ceramic hydroxyapatite Type II 40 µm 3 BioRad Macro-Prep ceramic hydroxyapatite Type I 80 µm 4 BioRad Macro-Prep ceramic hydroxyapatite Type II 80 µm 5 BioRad Macro-Prep ceramic fluoroapatite Type I 40 µm 6 BioRad Macro-Prep ceramic fluoroapatite Type II 40 µm 7 BioRad Macro-Prep ceramic hydroxyapatite Type I 20 µm 8 BioRad Macro-Prep ceramic hydroxyapatite Type II 20 µm 6

7 media MiniColumn Pseudo-Chromatogram for step elution (calculated by curve fitting) Optical density (540 nm) Optical density (540 nm) 0,5 0,4 0,3 0,2 0,1 0,0 0,6 0,5 0,4 0,3 0,2 0,1 0,0 Sample: 70 μl Vitamin B 12 in 1 M NaCl Eluant: 0.1 M NaCl Fraction size: 50 μl Elution Volume (μl) Superimposition of pseudo-chromatograms for 20 out of 96 columns Elution Volume (μl) MediaScout PipetColumn MediaScout PipetColumns are of 5 mm internal diameter and up to 10 mm bed height.they contain each up to 0.2 ml of the chromatography resins, packed to defined density, adjusted for each individual resin material, in order to replicate conditions in larger columns. Columns are run stepwise, while the flow is controlled by a positive displacement pipette and are designed for resin screening and for the early stages of method scouting. The volume applied can be up to several ml depending on the type of pipette used. MiniColumn efficiency typically replicates that of larger columns packed according to manufacturer s recommendation. Using a positive liquid displacement pipette: Set pipette into dispense mode. Regulate the speed to about 0.5 ml per minute Pipettes which work with our PipetColumns: Brand Handy Step Eppendorf Xstream (preferred for excellent low flow control) Gilson Repetman MediaScout CentriColumn MediaScout CentriColumns are of 5 mm internal diameter and up to 10 mm bed height. They each contain up to 0.2 ml of the chromatography resins, packed to defined density, adjusted for each individual resin material, in order to replicate conditions in larger columns. The liquid reservoir is 230 µl with 200 µl resin bed 280 µl with 100 µl resin bed 315 µl with 50 µl resin bed Columns are run stepwise, and are designed for resin screening and for the early stages of method scouting. MiniColumn efficiency typically replicates that of larger columns packed according to manufacturer s recommendation. Centrifuge working with CentriColumns: Hettich ROTANTA 460 RS or any 96 DIN centrifuge able to accept 110 mm high stack including base collection plate. Elution volume Peak half width Bed height/ column volume Theoretical plates per cm Asymmetry factor 214 µl ± 3,2 98 µl ± 4,1 1,1cm/216µl 24 ± 2,2 1,2 ± 0,15 Comparison of HR5/50 Amersham Bioscience column vs MediaScout MiniColumn Separation of Ribonuclease A, Cytochrome C and Lysozyme on Toyopearl SP 650 M, sample load 1% of breakthrough capacity A V [CV] A280 A528 mini column 1 mini column 2 Cond. Cond. mini column Cond. [ms/cm] Straight lines: Step gradient continuous elution of the 5/50 column with Äkta system. Dotted lines: Step gradient elution data of 50 µl fractions with MediaScout CentriColumns Data by Juergen Hubbuch,Tim Schroeder, Research Institute Juelich, Germany 7

8 media MiniColumn Fortsetzung. MediaScout MiniColumn MediaScout RoboColumn High Throughput screening columns in a 96 array format for robotic liquid handling stations These arrays extend the MediaScout product line by introducing mini columns for parallel analytical and preparative chromatographic separations with laboratory robotic workstations. These ready-to-use RoboColumns are available pre packed with any one from a large selection of commercially available chromatographic media. Up to 96 of them may be easily fixed onto a 96 well micro plate format, in any desired order. Comprising inner dimensions of 2.5 to 10 mm bed height and 5 mm inner diameter (c.v to 0.2 ml), the mini columns enable rapid parallel chromatographic separations with up to 96 columns, utilizing small sample and eluent volumes.allowing seamless cooperation between the RoboColumns and the robotic liquid delivery system, the column inlets are equipped with a patented needle adapter seal. This adapter permits the automated tight connection and disconnection of the robot needle unit to the mini column array. Chromatographic results may be gathered by normal detection methods, with individual fractions collected in standard micro plates. These MediaScout RoboColumns are 5 mm internal diameter and up to 10 mm bed height. They each contain up to 0.2 ml of the chromatography resins, packed to defined density, corrected for each individual resin material, in order to replicate conditions in larger columns. Longer RoboColumns up to 50 mm bed height are under development. Columns are run in principle stepwise, but could be run continuously when flow is controlled by a positive displacement system and applied by a fixed tip needle through a special needle adaptor.they are designed for extensive resin screening and for the early stages of method scouting in an advanced mode.the volume to be applied can be several ml depending on the type of system used. MiniColumn efficiency typically replicates that of larger columns packed according to manufacturer s recommendation. Workstations that can operate our columns: TECAN EVO-line other manufacturers, please check directly with Atoll 8

9 media MiniColumn Inner dimensions Bed volume Packing Flow rate Working temperature range 5-60 C Storage conditions Materials Filter plates Chemical stability 5 mm x mm (diameter x length) ml (to order) Technical Information: For all MiniColumn types: CentriColumn, PipetColumn, RoboColumn MiniColumns are ready-to-use and may be rearranged on the base plate in any configuration, according to the requirements of the proposed resin screening or methods development experiment. MediaScout MiniColumn arrays are delivered with capping mats and equilibrated in 20 % ethanol, with or without 0.15 M sodium chloride, depending on the particular resin.the array may be resealed with the cap mat after use and stored under appropriate conditions, e.g. at room temperature and equilibration with 20 % ethanol. Column position may be rearranged by the user should the need arise. Individual column rows may also be ordered separately for replacement purposes. Defined compression of chromatography media set to manufacturers specifications up to 5 ml/min (25 cm/min, 1500 cm/h) (depending on chromatography media) Equilibrated with 20 % ethanol (ion exchangers with the addition of 150 mm sodium chloride) polypropylene (column body, filter holder and plates) polypropylene/polyethylene 7 µm pore size ph 1-14, organic solvents (except for halogenated), mineral and organic salts, usual eluents and additives (urea, antioxidants, thiols, detergents) Note: Resin stability must be also considered Ordering information: For easy and convenient ordering of the MiniColumns, please visit to the SHOP section of our homepage Here you will find MiniColumn 12 row customizers. This customizer serves to help you configurating a 96-well setting with materials of your choice. Click on the MiniColumn version you want and fill the slots in the upcoming page with resins from the pull-down resin listing. Click on the Resin or on a Empty Slot to change it. Click on the topmost delete icon to empty all slots or on any other delete icon to empty the corresponding slot. After selecting the separation materials please print and fax this page to +49 (0) or save and this page to info@atoll-bio.com Price List To view up-to-date prices, please see (SUPPORT-DOWNLOADS) 9

10 media MiniChrom MediaScout MiniChrom: Useful formats and identical column hardware for all resin media types MiniChrom columns are delivered ready to use, packed with chromatography media from a large collection of commercially available resins or gel products for bioseparation. In the past no commercially available product allowed direct comparison of separation performances of dissimilar chromatographic media, with your own defined biological source material. Test kits from different manufacturers differ in design, volume of resin and geometry of test columns.as a result they were NOT directly comparable. Packing quality is variable and not directly comparable and reliable comparative performance data cannot reliably be achieved. MediaScout MiniChrom columns are primarily designed for use with chromatography systems.they may also be operated with syringes or peristaltic pumps.they may be reused as long as the resin remains in acceptable condition, which will depend largely upon the care taken, the cleaning performed and the tolerance of the resin to user conditions. Columns are not designed to be repacked by the end user. Features Ready-to-use LC columns Highly reproducible packing quality Benefits Greatly reduce development time and related costs Save tedious packing of columns Avoid packing skill variations between resin types User can focus attention on the separation task Delivers valid results Optimizes experimental procedures by selecting the best chromatographic media High quality small scale preparative purification Confidence in data replication between identical columns Available in 3 internal diameters: To make it easy to convert linear and volumetric flow rates. Internal Diameter (mm) Cross Section Area (cm 2 ) Volumetric flow (ml/min) Linear Flow (cm/min) Linear Flow (cm/h) 5, , , Available in versatile sizes: (mm.id.x mml) Crossover geometries: it is possible to choose alternative geometries for 1ml and 5 ml columns. 5 x 10 = 0.2 ml 5 x 25 = 0.5 ml 5 x 50 = 1.0 ml 8 x 20 = 1.0 ml 8 x 50 = 2.5 ml 8 x 100 = 5.0 ml 11.3 x 50 = 5.0 ml 11.3 x 100 = 10.0 ml 10

11 media MiniChrom Generate your own RESIN LIBRARY A truly innovative tool for the biochromatographer Using the Atoll MediaScout MiniChrom columns, to establish a chosen library of resins for the own particular applications allows customers to focus on value creation in bioseparation. Maybe you only need ion exchangers, maybe you need a range of chromatography modes, maybe you simply want to compare different particle sizes for a few resins Convenient As comprehensive as you wish to make it Direct comparisons between resins Low cost and High value Only Atoll GmbH can offer the following advantages YOU choose the resins for the library YOU choose the most suitable column size to suit your own needs From ANY of the available manufacturers Packed for highest performance (with optimised compression) In the SAME COLUMN HARDWARE for all media types For use on your own LC or HPLC system/workstation Very High Reproducibility Superimposition of HETP curves from 25 units out of a series of 200 MiniChrom mau Time (min.) 11

12 media MiniChrom MediaScout COLUMN GEOMETRY SELECTION GUIDE 5-10/0.2 ml This 5 x 10 = 0.2 ml column is designed to allow direct performance evaluations of the standard Atoll MiniColumn 96 array format units on a standard LC system, due to the use of standard connectors. This column is designed for this purpose alone. It can also be used in very small scale applications, e.g. when continuous monitoring is required. 5-25/0.5 ml 5-50/1.0 ml These two columns offer the smallest alternative volume units for traditional use on LC systems.when packed with medium to large particles (>20 µm) they can be expected to show qualitative differences between resin types of similar particle size, but due to inevitable wall effects cannot be expected to provide reliable quantitative data for predictive scale up calculations. 50 mm may be insufficient length for separation modes requiring higher plate numbers such as HIC, RPC and mixed mode resins. 8-20/1.0 ml This 8 mm ID unit offers a 1ml column for direct comparison with the 1ml 5 mm ID column. Bed height is shorter, but wall effects are less evident. Bed height will be unsuitable for valid evaluation of larger particle resins. It may however be useful when working with small valuable load samples and smaller resin particles, maybe as a final polishing step. This short column is not recommended for general purpose resin comparison with medium to large particles. This offers the closest comparison to a 1ml HiTrap column, but is available with any resin of your choice.atoll columns offer mid-range pressure resistance (up to 50 bar), when used in HPLC-systems. (HiTrap is a registered trade mark of GEHealthcare AB) 8-50/2.5 ml 8-100/5 ml These 8 mm ID 2.5 ml and 5.0 ml columns offer the best small scale comparisons for preparative and process resins. Choice of bed volume will depend on sample availability and value, and cost of resin.the diameter provides no significant wall effect. These units offer the smallest practicable model for general purpose comparisons, being the best compromise between column volume and efficiency. Smaller diameter units yield somewhat less reliable values for dynamic capacity and scale up resolution.at least 50 mm bed height is required for large particles. The longer, 100 mm column is recommended for separation modes requiring plate numbers for effective separations, such as HIC, RPC or mixed mode applications /5 ml /5 ml These 8 mm ID and 11.3 mm ID columns have the same 5 ml column volume, and permit evaluation of the effect of column bed height with identical volume units. These columns offer a small scale modelling of dynamic performance when transferring methods from linear gradient to stepwise gradient conditions, and to ensure method robustness when working up step elution methods /5 ml /10ml The 11.3 mm ID geometry provides a convenient 1cml/ml format for easy scale up calculations involving flow rate, dynamic capacity and resolution, etc. Volumetric flow in ml/min numerically equals linear flow rate (cm/min). These columns offer a scale down model suitable for predictive scale up calculations. The longer, 100 mm column is recommended for separation modes requiring plate numbers for effective separations, such as HIC, RPC or mixed mode applications. Methods may be transferred directly to Atoll ValiChrom glass columns which offer an identical diameter with a fixed, custom bed height up to 60 cml. ValiChrom prices are available on the Atoll website 12

13 media MiniChrom Packing Storage medium Flow rate Connections Maximum working pressure Temperature ranges Chemical Stability of column hardware Chromatographic performance Technical Information: MediaScout MiniChrom is a miniaturized low to medium pressure chromatography column format, used in bioseparation applications, accommodating small sample volumes. Components of MediaScout MiniChrom columns are made from biocompatible polymers, mainly polypropylene (column tube, end piece, filter adapter). Filter plates are made from porous polypropylene material. Columns may be connected directly to most liquid chromatography systems or workstations and connection to earlier FPLC systems is simple with standard male / M6 female adapters. Unless otherwise stated, packed media are equilibrated with 20 % ethanol or a mixture of 20 % ethanol and 150 mm sodium chloride (ion exchangers, IMAC and HIC resins). Columns are equipped with plastic fittings and may be stored at room temperature. Used with chromatographic media of all types with particle size > 10 µm Defined compression of chromatographic materials defined by manufacturers specifications or customized 20 % Ethanol with or without 150 mm NaCl, or customized Up to 10 ml/mino Standard HPLC (M10-32 UNF for 1/16 tubing) 30 bar During separation: -10 C +60 C, storage: 10 C 30 C ph 1-14, organic solvents (except halogenated), minerals and salts, common eluents and additives (urea, antioxidants, thiols, detergents, tensides) In accordance to separation material and the column geometry (I.D. and bed height) and compression (e.g. 2,500 tp/m for Toyopearl 650 (M)) Ordering Information: Packs of 5 x prepacked columns, off the shelf or according to your specifications enable a direct comparison of resin performance. Choose your specific settings and media in a kit, Atoll will deliver them prepacked in columns, readily configured to meet your needs. Customized media packing is also available For easy ordering of the MiniColumns, please visit to the SHOP section of our homepage Here you will find MiniChrom customizers for the different diameters. This customizer serves to help you to order columns with materials of your choice. Click on the MiniChrom version you want and fill the slots in the page with resins from the pull-down resin listing. After selecting the separation materials please print and fax this page to +49 (0) or save and this page to info@atoll-bio.com Price List To view up-to-date prices, please see (SUPPORT-DOWNLOADS) 13

14 media ValiChrom MediaScout ValiChrom: Precision medium pressure range glass column system for parameter evaluation and validation. cgmp Validation columns of 1cm 2 cross section area and precisely to the length of customers DSP column for scale-down experiments in lot-to-lot evaluation, CIP, virus clearance, etc. Features Professionally packed Documented supplied mm I.D. = 1ml/cm geometry, any length, up to 600 mm Unused or used resin provided by user cgmp manufactured and sterile packing on request Benefits Low cost, convenience and quality assured The ValiChrom represent a scaledown model of the respective full-scale column Ideally suited for parameter evaluation and optimisation User defined resin for validation studies For immediate use in validation studies MediaScout ValiChrom glass columns are available customized to different inner geometries and are professionally packed with any resin or chromatography media. These are individually flow-packed, either according to resin manufacturers recommendations or customized to model conditions in corresponding full scale columns. Thus, the ValiChrom represent a valid scale-down model of the respective fullscale column and are suited for parameter evaluation and optimisation, as well as for process validation purposes, utilizing the original bed height and resin compression at the smallest practicable inner diameter. The ValiChrom range is designed for qualitative and quantitative resin screening experiments. ValiChrom columns can also provide an accurate, valid model using smallest sample size volume for small scale methods development work.they are unique in their ability to provide a reliably packed and valid evaluation system for the full range of resin types. MediaScout ValiChrom columns are primarily designed for use with chromatography systems. They may be reused as long as the resin remains in acceptable condition, which will depend largely upon the care taken, the cleaning performed and the tolerance of the resin to user conditions. ValiChrom columns can be refilled by Atoll. You can specify prepacked columns to perform your validation of lot-to lot consistency of resins or for virus and other validation studies, (e.g. CIP or clearance). Professionally packed into precision bore glass columns with accurate compression to represent conditions in your production column for a price that cannot be matched. Atoll will pack any resin of your choice or your own specific resin into columns with the length to match the purification scheme of the customer. Validation columns 1ml/cm l Lengths from cm Certified with asymmetry factor, plate number and pressure flow data. ValiChrom columns are available in inner diameters of 11.3 and on request 16 mm and 25mm with any fixed bed heights ranging from 10 up to 60 cm. The simple fixed bed height design of the ValiChrom columns avoids complicated adaptor construction.therefore, these packed columns are available at very competitive prices. 14

15 mediascout Column Type Inner Diameter x Length (mm x mm) Column Cross Section cm 2 Technical Information: MediaScout MiniChrom ValiChrom columns are shipped ready to use, packed with chromatography media from the large range of commercially available resins or gel products for bioseparation. Unless otherwise stated, packed media are equilibrated with 20 % ethanol or a mixture of 20 % ethanol and 150 mm sodium Column Volume ml Flowrate1 ml/min / cm/h Max. Flowrate 1 ml/min / cm/h ValiChrom 5 4 -X 5.0 x / 150 < 5 / 1500 ValiChrom 8 4 -X 8.0 x / 150 < 5 / 1500 ValiChrom X 11.3 x / 150 < 25 / 1500 ValiChrom X 16.0 x / 150 < 25 / 1500 Connectors Maximum Operating Pressure 1 Chemical Stability 2 standard HPLC (M10-32 UNF for 1/16 inch capillaries) 30 bar Columns are tolerant to aqueous buffers and salt solutions, 1 M alkaline solutions, 1 M non-oxidizing mineral acids, 8 M urea, 6 M guanidine hydrochloride, non-halogenated organic solvents and detergents.they are not compatible with strong oxidants and halogenated solvents 1 This depends on the chromatography material. 2 The chemical stability refers to the column hardware parts only.the individual packed chromatography media may have different stability. To avoid damage to the chromatography media, please refer to the relevant manufacturer s data. 3 The precise ID of the glass tube is mm, corresponding to 1cm 2 cross section. 4 In preparation. chloride (ion exchangers, IMAC and HIC resins). Columns are equipped with plastic fittings and may be stored at room temperature. Components of MediaScout ValiChrom columns are made from glass (column tube) and biocompatible polymers, mainly highdensity polyethylene or polyoxymethylene (column tube, end piece, filter adapter). Filter plates are made from porous polypropylene/polyethylene material.the protective outer tube is made from acrylic glass. Columns may be connected by standard HPLC/ÄKTA connectors directly to most liquid chromatography systems or workstations and connection to earlier FPLC systems is simple with standard male / M6 female adapters. Ordering Information: ValiChrom cgpm Glass columns are individually quoted due to a large variety of resins and column geometries. Resin can be provided by customer if necessary (Lot specific etc.) Please specify your resin and the column dimensions to info@atoll-bio.com Price List To view up-to-date prices, please see (SUPPORT-DOWNLOADS) 15

16 media MaxiChrom MediaScout MaxiChrom: cgmp, disposable/incinerable columns The universal, disposable column system for preparative applications and scale-up development. MediaScout MaxiChrom columns are delivered ready to use, professionally packed with chromatography media from the full range of commercially available resins or gel products for bioseparation. E.g. 100-X columns (X = mm,) are packed with any resin or chromatography media chosen by the user, preferably with materials of particle size larger than 50 µm. Features cgmp, disposable, fully incinerable production columns Ideal as guard-columns to protect valuable capture resins Produced according cgmp standard, low bioburdon, process suitable connectors 32mm, 40mm and 100mm i.d. Customer defined bed height User defined resin They are individually flow-packed to take account of the varying compressibility of each resin. Bed heights are fixed to an accuracy of ± 1mm. MaxiChrom 100-X chromatography columns are designed for preparative applications and/or scale-up development work. The column hardware is fully incinerable, which makes the columns particularly useful for single use in bio-pharmaceutical production. MediaScout MaxiChrom columns may be reused as long as the resin remains in Benefits For situations when validation of contaminant removal is critical Avoids dismantling and repacking of the production column Immediately useable in the production train Pilot scale and production sizes Custom designed clumns acceptable condition, which will depend largely upon the care taken, the cleaning performed and the tolerance of the resin to user conditions.the columns are not designed to be repacked.the column hardware can be completely incinerated. Sterilization of packed columns by autoclaving is possible with many of the commercially available sorbents. Columns for special applications Packed with adsorbants as disposable (incinerable) device for removal of detergents after virus inactivation during production. Custom packed separation media for virus removal during production. Packed separation media for virus validation experiments during registration of a separation scheme in completely incinerable columns. Packed as process scale guard columns to protect high value media in process capture columns. Packed with any resin for flow-through polishing to remove specific residual contaminats. 16

17 media MaxiChrom Technical Information: Unless otherwise requested, packed media are equilibrated with 20 % ethanol or a mixture of 20 % ethanol and 150 mm sodium chloride (ion exchangers, IMAC and HIC resins). Columns are sealed with threaded end caps, including a TriClamp 1/2 gasket and may be stored at room temperature. Components of MediaScout MaxiChrom columns are made from bio-compatible polymers. End pieces and filter adapters are from polyoxymethylene (POM), column tubes from acrylic glass and O-rings are from Viton. Frits and flow distributors are made from polypropylene. Columns may be connected directly to, and operated by, normal liquid delivery systems (e.g. gear or peristaltic pumps) or chromatography workstations, using standard quarter inch Mini TriClamp fittings to make connections between liquid delivery system and column. It is recommended to continuously measure the operating pressure drop by connecting a pressure gauge in-line between pump and column. Ordering Information: MaxiChrom cgpm disposable/ incinerable columns are individually quoted due to a large variety of resins and column geometries. Please specify your resin and the column dimensions to info@atoll-bio.com During 2007/2008 Atoll expects to expand this range with larger diameter MaxiChrom disposable units. Price List To view up-to-date prices, please see (SUPPORT-DOWNLOADS) Properties of MediaScout MaxiChrom 100-X columns (two examples with different bed heights outlined). Column Type Resin packing Connectors standard Mini TriClamp 1/2 Performance Pressure maximum Temperature range Storage conditions Materials Filter plates (frits) Chemical stability 2 Inner Diameter x Length (mm x mm) flow packing to defined compression, according to resin specification depending on chromatographic medium 1.5 bar Column Cross Section cm 2 Column Volume ml Operation/Storage: 5 C - 60 C / 5 C - 30 C Flowrate1 ml/min / cm/h equilibrated with 20 % ethanol (ion exchangers with additional 150 mm sodium chloride) Polyoxymethylene (POM) (column tube, filter adaptor, end piece, end cap), Viton (o-rings) Polypropylene/polyethylene (17 µm pore size, others on inquiry) Max. Flowrate 1 ml/min / cm/h MaxiChrom x / 229 < 800 / 611 MaxiChrom x / 229 < 800 / 611 Columns are tolerant to aqueous buffers and salt solutions, ph 2-14, 1 M alkaline solutions, 1 M non-oxidizing mineral acids, 8 M urea, 6 M guanidine hydrochloride, ethanol, isopropanol and detergents 1.They are not compatible with strong oxidants and most organic solvents 1 This depends on the chromatography material. 2 The chemical stability refers to the column hardware parts only.the individual packed chromatography media may have different stability. To avoid damage to the chromatography media, please refer to the relevant manufacturer s data. 17

18 media ResiQuot MediaScout ResiQuot: for a accurate and reproducible dispensation of aliquots The MediaScout ResiQuot is a vacuum manifold device for accurate and reproducible metering of aliquots from solids or suspensions such as chromatography media ( Resin aliquots ), into standard 96-well microplates. MediaScout ResiQuot is a vacuum manifold device for the accurate and reproducible preparation of aliquots from solids of suspensions, e.g. from chromatography media, into any kind of 96-well micro plates. Subsequently, these are used in experimental determination of chromatographic parameters in batch mode and other applications. Applications for batch tests in 96-well microtiter plates. To obtain quick and convenient adsorption isotherm data. Optimization of adsorption conditions. Fast performance comparison of separation media. Drastic reduction of development time and cost. MediaScout ResiQuot functions with very small amounts of chromatography media or other solids available in a suspended form. The smallest useful sizes of aliquots contain a volume of as little as 7.8 µl ± 0.3 µl, other available volumes are 20.8 and 50.9 µl. Aliquot preparation using pipettes does not generally allow such a high accuracy, except when strongly diluted suspensions are used.the latter method is associated with additional time consuming procedures to remove the residual liquid after aliquotation. Features Generates 96 precison aliquots Speedy preparation of aliquots For all kind of slurry suspensions Benefits Enables accurate metering of solids in suspension, e.g. separation media. Aliquot preparation by pipette does not generally allow such a high degree of accuracy, even with highly diluted suspensions due to tip blocking and sedimentation etc. Avoids time-consuming removal of residual liquid after aliquot dispensation Materials displaying a tendency to stick to surfaces or to settle rapidly are particularly difficult and unreliable to sample by pipette, even from highly diluted suspensions. Precision volumes of resins of all types and particle size can be prepared 18

19 media ResiQuot A Materials displaying a tendency to stick to surfaces or settling rapidly are particularly difficult to meter acurately and are unreliably pipetted, even from highly diluted suspension B (A) Sketch of a complete MediaScout ResiQuot unit and a unit with upper part enclosure removed (B), including parts as indicated: upper part enclosure (1) knurled head screws (2), bottom part enclosure (3), clamping plates (4), stopper plate (5), slider (6), vacuum connector (8), gasket (9), perforated plaque plate (10) frit holder with stainless steel frit (10 µm) (11). The functional principle of the MediaScout ResiQuot, is to aspirate the solid particles from a homogeneous suspension in a well defined consistant manner into the 96 cylindrical holes of a metal plate, the perforated plaque plate. Small cylinders of aspirated solids are released from these holes using a pusher tool.this metal tool comprises 96 cylindrical spikes corresponding to the holes in the plaque plate, which push the aliquots directly into the 96 wells of a microplate placed underneath. Excess solids remaining in the holes of the upper part of the device can be retrieved similarly and used for the preparation of more aliquots MediaScout ResiQuot Technical Details and Function The lower plate (3) of the device functions as a manifold chamber with vacuum connector and frit holder (11) with stainless steel frit. The edge of the frit holder is completely sealed against the lower plate (3) by the gasket (9). Vacuum aspiration can only take place through the stainless steel frit. The precise alignment and orientation of the upper manifold (1) onto the lower manifold (3) is assured by two dowel pins placed near the edge of each unit and fitting into drilled holes. Two knurled assembly screws (2) provide rapid and secure clamping of the upper and lower manifolds. A gasket inserted into and around the edge of the upper manifold ensures a vacuum tight connection between upper and lower manifolds. Once assembled, the perforated plate is inserted into the front opening as far as to the stop. The 96 holes drilled in the perforated plate are then aligned to the centres of 96 larger holes in the upper manifold. A vertical metal plate acts as a stop to ensure precise alignment and includes a gasket seal between the perforated plate and the assembled manifolds. The perforated plate is held firmly in place by two clamps (4), attached to the front of the upper manifold. The holes drilled in the upper manifold are filled with the required suspension which is then drawn into the 96 small cylindrical holes of the perforated plate under vacuum, to the level of the stainless steel frit. 19

20 media ResiQuot As long as there is sufficient solid material in the suspension the cylindrical holes of the perforated plate are evenly and completely filled. Excess solids remaining in the drilled holes of the upper manifold may be recovered after removal of the sample aliquots. The perforated plate is released and withdrawn over wipers, which remove any residual solids from the plate surface. Compacted solid material is retained in the precisely filled cylindrical holes of the perforated plate. The upper and lower manifolds are dissembled and the upper unit cleared of excess material using the 8-way pusher tool. In this way, the unit may be used to sample a single suspension into 96 separate wells, or each well may be filled separately with materials of different composition depending on the needs of the user. Technical Information: MediaScout ResiQuot Dimensions (enclosure only) Total outer dimensions of mounted device Weight mounted device including perforated plate 96-fold aliquot removal tool for perforated plate excess material recovery tool 8-fold for upper part enclosure Format Aliquot volume Materials Enclosure Gaskets 16/12/7.6 (L/D/H cm) 17/19.5/9.2 (L/D/H cm) 2.8 kg 394 g 76 g Compatible with 96-well micro plates (according to ANSI standard) 7.8 µl (smallest available dimension), 20.8 and 50.9 µl available on request Anodically oxidized Duralumin Silicone rubber Frit Stainless steel 0.07 mm thick, pore size 10 µm Screws stainless steel Chemical stability (at R.T.) Aqueous solutions in the range of ph 4 to 12 Pressure stability Vacuum of 1 bar Figure 3. Example of 96 identical aliquots pushed into a 96-well plate. The perforated plate is then placed above a 96 well collection microplate, and the compacted solids in the cylindrical holes are transferred into the wells using the 96 well format removal tool. The pusher tool is precisely aligned to the 96 cylindrical holes by four guide pins located at the corners of the tool. An example of resin aliquots obtained by transfer from the perforated plate into a microplate is shown in figure 3. Finally, excess solids are recovered from the uppermanifold. Ordering Information: ResiQuot units are individually quoted due to individual requirements of resin aliquotes. Please specify your required volume slider and push-out unit to info@atoll-bio.com Price List To view up-to-date prices, please see (SUPPORT-DOWNLOADS) 20

21 mediascout Frequently asked questions 1. What is the best method of using the MiniColumns? k Three different functional designs are available a) CentriColumn: Manual or robotic pipetting of liquid (sample and eluants): Columns integrate a reservoir located on top of the resin which holds samples or eluants. Flow through the column is induced by centrifugation. (Abt rpm, 1 min.) Results are obtained by sequential pipetting, centrifugation and plate reading. b) PipetColumn: Transport of liquid by a positive liquid displacement (pld) pipette, (preferable motorized). Instead of the reservoir on top of the resin, there is a patented adaptor that seals with most of the common pld-pipette tips. Positioning the tip on top of the column, holding by gentle pressure and moving the liquid by setting the slowest dispense rate. In this way all columns are operated at the same linear flow rate. Volumetric flow is accurately controlled during every pass through the column. Eluate can be quantitatively collected ensuring accuracy between columns. Droplets coming out from the column will vary according to viscosity of the mobile phase but is in the range of µl. The StarterKit, which contains one row of 8 Pipet- Columns can deliver 12 different elution conditions in parallel by shifting the 8-row one position further after each elution step. c) RoboColumn: These columns also have the same geometry as MiniColumns and can be operated by robotic flow delivery systems. Instead of the reservoir on top of the resin, a needle adapter with a patented sealing system allows you to operate the column as in an LC system. Droplets coming out from the column have usually a volume of 28 µl. NB: MiniColumns are NOT designed to work with vacuum for liquid transport through the resin, which can dry the column and destroy the integrity of the packed bed. In addition, the depth filters used in MiniColumns, cause no significant flow resistance. In combination with different resin flow characteristics, this would potentially cause one of the 96 columns to dry out first and all the air would be sucked through that column. So in the 95 others the liquid would not move further.the same would happen using positive air pressure to move liquids. Applying vacuum to samples containing protein, can cause tremendous foaming at the lower column outlet which leads to cross contamination all over the plate. 2. Which resins are available in MiniColumns? k Atoll offer a wide range of resins for column packing this can be viewed on the website. Or customers can provide their own resin which Atoll will pack.this enables Atoll to remove resin supply costs for the more expensive resin types and to offer resin packing for custom special resin. Please refer to our resin listing on 3. How are separation materials packed into MiniColumns and MiniChrom? k Atoll applies flow packing to obtain a defined packing density, similar to large scale columns. This results in packing quality similar to that of well packed large process columns. 4. Can MiniColumns or MiniChrom units be refilled? k No, once a prepacked column is dismantled, the housing is damaged and no re-use possible. However the column can be regenerated as many times as the respective resin allows. 5. Are there prepacked columns available with rigid materials like silica or hydroxyl apatite or ceramics? k Yes, materials like silica, hydroxyl apatite or ceramics are available in both systems.the special inlet adapter in MiniChrom allows Atoll to pack all material types with high efficiency. 6. What are the dimensions of MiniColumns, MiniChrom and ValiChrom? k a) MiniColumn: Polypropylene columns, precision molded, all are 5 mm ID. Max volume is 200 µl, (used for HIC and IEC materials) other volumes of 100 µl or 50 µl, used for high priced resins like ProteinA materials are available on request. For SEC mechanisms, in particular for Desalting the columns need to be longer in bedheight to deliver baseline or acceptable separations. For special purpose of desalting relative large volumes in robotic handling stations, MiniChrom barrels are available, mounted on a fitting 96array plate, equipped with a MediaScout needle port to be run in fixed tip robotic stations: Examples for such special larger units 24 units 5-50 mm 24 units 8-50 mm 24 units mm 16 units mm 16 units mm b) MiniChrom: Polypropylene columns, precision molded 5 mm ID - 10 mm length = 0.2 ml 5 mm ID - 25 mm length = 0.5 ml 5 mm ID - 50 mm length = 1.0 ml 8 mm ID - 20 mm length = 1.0 ml 8 mm ID - 50 mm length = 2.5 ml 8 mm ID mm length = 5.0 ml 11.3 mm ID - 50 mm length = 5.0 ml 11.3 mm ID mm length = 10.0 ml b) ValiChrom: Glass columns, precision bore (Schott), PMMA protection jacket mm ID mm length = ml 7. Which chromatography system is suitable for the operation of MiniChrom? k Any HPLC- or mid- or low pressure system, capable of delivering accurate flow rates of 0.1ml/min and higher. Furthermore any other liquid delivery system, e.g. a peristaltic pump or a syringe can be used with an appropriate adapter. 8. What is the backpressure of a 5-50 MiniChrom column? k Depending on the resin, backpressure ranges between 0.1 bar up to 10 bar at 150 cm/h. Usually when operating 40 µm resins, backpressure is 1-2 bar at flow rates of 600 cm/h. 9. Why is ValiChrom relatively inexpensive compared to other columns of that kind? k ValiChrom columns are constructed from precision bore glass (Schott), protection jackets from acrylic glass and end pieces machined from POM or PP, avoiding complicated and cost intensive adjustment systems.the desired length of the resin bed is precisely reached by a standard bottom adaptor and a top adaptor machined to achieve the customer desired bedheight together with a fitting glass tube length by +/- 1mm 10. What is the purpose of MaxiChrom columns? k MaxiChrom columns are designed for use in production of biomolecules, when validation of contaminant removal is critical. Examples are production of virus probes or removal of detergents like Triton 100 with reversed phase resins after virus inactivation in bioprocessing schemes. MaxiChrom columns are usually packed with resins of >100 µm particle size and have therefore relatively low back pressure values. As standard version, they consist of acrylic glass barrels and POM endpieces, sealings are Viton, frits and distribution plates are PP. Guard columns (e.g. for Protein A resins) Low cost disposable polishing process columns 21

22 U A m injection collect 30 m A20 U 10 injection collect Time (min) Applications mediascout Validation Columns MediaScout ValiChrom scale-down columns Method development Check product retention on resin Determine Separation Condition (resins, ph, salt type, etc ) Binding Experiments on Mediascout MiniColumns Capture step Intermediate step with 1cm 2 inner ID, and exact identical length to customer s DSP column, up to 600mm long Convenient handling 96 array plate with MiniColumns 1.Bind apply sample by pipette or robot 2.Wash / Elute move liquid by centrifuge or pld / needle-tip 3. Collect 4. Analysis 96 Pseudo Chromatograms in <1 hour Bioreactor Polishing step Reproducibly packed with customer s resins for all kind of validation issues: Top filter plate Bottom filter plate Sa mp le c ha mb e r Re sin c h a m b er Column outlet (Luer) Collect eluate Established DSP- Purification Scheme lot consistency cleaning virus removal column : 450 µl resin : 200 µl Liquid : 240 µl Sample : ~400 µg UV & HPLC or SELDI-TOF High-Throughput Screening (HTS) with mediascout MiniColumns Determination of maximum salt concentration for HIC binding (example) on the same resin in each MiniColumn. High-Throughput Screening (HTS) with mediascout MiniColumns To find the right Resin and Eluant Condition Determine Separation Condition (resins, ph, salt type, etc ) 6 salt types 4 salt concentrations 4 ph levels & 4 sample concentrations in each column (from the lowest sample concentration to the highest) Add sample onto HIC resin in high salt Adding solution with lower salt concentration at each stage => centrifuge => collect eluant => read collector plate => etc. 384 conditions evaluated Bioreactor Monitoring Desalting in 1 Minute on 0.5ml columns Capture step 1. apply 2. wash Take aliquots of clear bioreactor supernatant Apply to MiniColumn* packed with the same capture resin that is used in the DSP scheme Target molecule is bound Typical HPLC chromatogram of protein desalting using MediaScout MiniChrom 5-25, packed with Sephadex G25 Superfine, 20µl sample, 1mg/ml Ovalbumin, 10mg/ml NaNO 3, flow rate: 150 cm/h This degree of desalting is sufficient for MS applications and can be transferred to automation on 96-array format! Ovalbumin NaNO 3 Intermediate step impurities 90% bulk impurities pass through 3. elute Bioreactor Polishing step target molecule Submit to high resolution HPLC, or similar, to 4. check check product pattern Established Purification Scheme * CentriColumn and PipetColumn for manual handling, RoboColumn for automated procedures In-Process-Control e.g: mab Bioreactor Monitoring In-process control in fermentation research Fermentation supernatant, e.g. antibodies in mammalian cells, conc µg/ml Fermentation broth, e.g. antibodies in mammalian cells apply waste wash Adsorption of mab on Protein A MediaScout RoboColumn in 96well format Filtration at 0.45µ to remove cell debris elute directly apply 50 µg to IE-HPLC Desalting on MediaScout RoboColumn In 96-well format directly apply to Mass Spectroscopy Desalting in 2 Minutes on 5ml columns Typical HPLC chromatogram of protein desalting using MediaScout MiniChrom 8-100, packed with Sephadex G25 Fine, 500µl sample(10%vc), 1mg/ml BSA, 10mg/ml NaNO 3, flow rate: 150 cm/h BSA 200 NaNO Time (min) 22

23 Ettishofer Straße Weingarten Telefon +49 (0) Telefax +49 (0) info@atoll-bio.com Internet: mediascout