Evolve 250 Technical User Manual
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Evolve 250 Technical User Manual

70mm, 100mm and 140mm chromatography columns
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70mm, 100mm and 140mm
Chromatography Columns

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Summary of Contents for Evolve 250

  • Page 1 70mm, 100mm and 140mm Chromatography Columns...
  • Page 2 DELTA PRECISION LTD Chromatography Column Operating Manual and Installation Guide 70mm, 100mm and 140mm Columns...
  • Page 3 Technical specification updated caster not available on 70-140mm columns 22/04/2021 Updated to reflect Evolve® Trademark status 12/07/2021 Evolve® is a registered trademark of Delta Precision Ltd. Evolve® is a registered trademark in the EU, UK and US. Manual Revision Date...

  • Page 4: Table Of Contents

    Kits, Spare parts and Accessories ................. 29 Spare parts and kits ..........................31 Spare Parts and kits Cont........................32 Accessories ............................34 Contact Us ........................35 Appendix 1 Chromatography Performance ..............36 Performance Measurement and Column Efficiency ................. 36 Performance Results From Evolve®Columns ..................38...
  • Page 5 T able of Figures Figure 1.1 Evolve®Column Range 70mm, 100mm, 140mm, 200mm and 250mm columns ....2 Figure 1.2. Pressure Drop Evolve®70mm column ..................4 Figure 1.3 Pressure Drop Evolve®100mm column ................... 4 Figure 1.4 Pressure Drop Evolve®140mm column ................... 5 Figure 3.1 Diagram showing the location of the fixed bed support .............

  • Page 6: Column Hardware

    • Prevents any product inactivation through metal contact Unlike other columns available on the market the Evolve®columns have an integral bed support and seal assembly. The polypropylene mesh and seal are manufactured as a single unit and fits directly on the flowcell without any fixings.

  • Page 7: Figure 1.1 Evolve®Column Range 70Mm, 100Mm, 140Mm, 200Mm And 250Mm Columns

    20-520mm. • Tube unit – consisting of the column tube, flanges and column stand or support legs • Fixed assembly – consisting of the bottom flowcell assembly and flowtube. Figure 1.1 Evolve®Column Range 70mm, 100mm, 140mm, 200mm and 250mm columns...

  • Page 8: Technical Data

    Maximum height mm 1523 1543 1545 1550 1550 Minimum 1620 1620 1620 1750 1750 Clearance** mm casters are not available on 70-140mm columns *based on bed height of 520mm and specific gravity 1.05, **minimum height required to remove adjuster from tube unit.

  • Page 9: Pressure Drop

    15-20 C using DI water. Volumetric Flowrate L/min Evolve 70 1000 1500 2000 2500 Linear Velocity cm/hr Figure 1.2. Pressure Drop Evolve®70mm column Volumetric flowrate L/min Evolve 100 1000 1200 Linear Velocity cm/hr Figure 1.3 Pressure Drop Evolve®100mm column...

  • Page 10: Figure 1.4 Pressure Drop Evolve®140Mm Column

    Volumetric flowrate L/min Evolve 140 1000 1500 2000 2500 3000 Linear Velocity cm/hr Figure 1.4 Pressure Drop Evolve®140mm column...

  • Page 11: Material Conformity

    Material Conformity All materials in contact with the process stream have been selected for their suitability for use in equipment utilised in Biopharmaceutical applications and so either conform to relevant sections of FDA code of Federal Regulations vol. 21 170-199 and/or passed USP class VI test for in vivo toxicity.

  • Page 12: Chemical Compatibility

    ¨ ¨ Dimethyl formamide ¨ ¨ Dimethyl sulfoxide 10% ¨ ¨ Disodium phosphate ¨ ¨ Ethanol 20% ¨ ¨ Ethanol 70% ¨ ¨ Ethylene glycol (1,2-ethanediol) ¨ ¨ Formaldehyde 50% ¨ ¨ Glycerol (Glycerine) ¨ ¨ Guanidine hydrochloride 6M ¨...
  • Page 13 Methyl chloride (Chloromethane) ¨ ¨ Methyl ethyl ketone (MEK) ¨ ¨ Methylene chloride ¨ ¨ Nitric acid 10% ¨ ¨ Nitric acid 70% ¨ ¨ Peracetic acid 300 ppm ¨ ¨ Phosphoric acid ¨< 55% ¨ Potassium hydroxide 2M ¨ ¨...

  • Page 14: Safety Guidelines

    The design and Manufacture of chromatography column including the Evolve®column are affected by these regulations. The regulations are divided into Categories and the Evolve® column has been assessed and found that it falls within the Sound Engineering Practice (SEP) category. The foundation of the assessment is based on the following;...

  • Page 15: Chemical Compatibility

    Larger Columns and their subassemblies are particularly heavy and will necessitate a ‘2 Person’ lift and the subsequent use of mechanical handling equipment or manual handling aids. Provision is made for the use of a hoist. It is essential that ‘good practice’ is adopted regarding the use of appropriate slings and associated equipment.

  • Page 16: Installation And Column Set-Up

    Column Preparation Installation of bed supports The Evolve® range of columns features polypropylene, woven mesh bed supports, integrated with a Santoprene seal. These do not require prewetting before use and are required to be fitted directly onto the adjuster and fixed flowcell.

  • Page 17: Figure 3.2 Photograph Showing The Installation Of The Fixed Bed Support

    Figure 3.2 Photograph showing the installation of the fixed bed support 3. Locate fixed flowcell into the tube unit and fix in place. Leveling the 70mm, 100mm & 140mm Columns After installation of the fixed flowcell and before installation of the adjuster unit, it is recommended to level the column.

  • Page 18: Figure 3.3 Diagram Showing The Location Of The Adjuster Bed Support

    Installation of Adjuster Bed Support 1. The adjuster assembly should be removed from tube unit and placed on its side on a clean work surface. 2. With the rough side facing towards the ribbed flowcell, press the edge of the seal into the location groove on the side of the flowcell.
  • Page 19 Set Bed Height 1. Reinstate the adjuster unit in the tube, using the location pins and ensure the adjuster seal is de-energized‘ 2. Replace the flange fixing bolts underneath the tube flange and tighten. 3. Partially energized the seal and turn the height adjust handles clockwise to set the required bed height.

  • Page 20: Column Operation

    Chapter Column Operation This section provides general guidance on operating the Evolve® range of columns. It includes a hydrostatic leak test to establish that the column is installed correctly and advice for the preparation of the chromatography media. Further advice specific to each media can be obtained from the media manufacturers; this section only provides a general guidance.

  • Page 21: Media Preparation

    Media Preparation Below provide a general method for preparing and packing the Evolve® columns. However it is advisable to follow the media manufacturer’s recommendation for preparation of the media. 1. The media should be prepared as a 50 – 75% slurry in equilibration buffer, packing buffer or other suitable solvent.

  • Page 22: Figure 4.1 Schematic Of The Column With Part Identification

    Figure 4.1 Schematic of the column with part identification 16. Pump buffer into the column at an appropriate rate to pack the bed. Ensure fixed end valve is open. The packing flowrate should be a minimum of 10% to 20% higher than the highest flowrate to be used during the process.

  • Page 23: Unpacking

    (refer to media manufacturers data). Common storage solutions are 0.1 - 0.5M sodium hydroxide or 0.1M sodium hydroxide + 20% ethanol, both of which are suitable for long term (several months) storage with Evolve® columns.
  • Page 24 Unpacked columns should be kept in a cool, dry, dust-free location and protected from light. In particular, do not expose acrylic column tubes to direct sunlight for prolonged periods or sterilize with ultra-violet lights as this may reduce the mechanical strength of the tube.

  • Page 25: Maintenance

    Chapter Maintenance Routine Maintenance Procedures-General guidelines To disassemble and assemble the columns follow the instructions below. After reassembling the column ensure that the column does not leak by performing a hydrostatic leakage test. When assembling/dissembling attention should be paid to: Always use the correct size tools.

  • Page 26: Disassembly Of 70Mm, 100Mm And 140Mm Columns

    Disassembly of 70mm, 100mm and 140mm Columns Adjuster Assembly 1. Detach any pipework from the top flowtube adapter. 2. Remove flange fixing bolts from the underside of the top tube flange. 3. Check that adjuster seal is released by turning the seal actuation nut counter clockwise. 4.

  • Page 27: Figure 5.1 Adjuster Assembly Evolve® 100 Column

    Figure 5.1 Adjuster Assembly Evolve® 100 column Fixed Assembly 1. Disconnect the pipework elbow from the bottom of the column. 2. Using the fixed end fixing tool and torque wrench, loosen each of the fixing screws, supporting the flowcell with your hand. Remove the flowcell and place it on a clean work surface.

  • Page 28: Figure 5.2 Fixed Assembly Evolve® 100 Column

    Figure 5.2 Fixed Assembly Evolve® 100 column Tube assembly The unit should only be disassembled as apart of an emergency repair procedure. Details of the method will be provided with the replacement parts.

  • Page 29: Reassembly Of 70Mm, 100Mm And 140 Column

    Reassembly of 70mm, 100mm and 140 column. Reassembly of the column is the reverse of disassembly. Fixed Assembly 1. Install the process flowtube into the back of the flowcell. Ensure the flowtube seal is located. Tighten process flowtube using correct tools. 2.

  • Page 30: Figure 5.4 Photograph Showing The Location Of The Adjuster Bed Support

    Adjuster Assembly 1. Install the adjuster mesh by carefully locating the bed support seal into its location groove in the side of the adjuster cell. Ensure the seal is pushed tight into the groove. The bed support should be fitted with the smooth side facing into the column. (Rougher side to flowcell).

  • Page 31: Figure 5.6 Photograph Showing The Final Location Of The Adjuster Bed Support

    4. Locate the flowtube within the stainless steel central seal setting tube and replace the flowcell locking clip. Adjuster Seal Seal Figure 5.6 Photograph showing the final location of the adjuster bed support 5. Locate the flowtube/central seal setting tube inside the threaded height adjust rod and refit the spacer and central adjuster nut.

  • Page 32: Troubleshooting

    Chapter Troubleshooting Refer to the maintenance section before undertaking any procedure advised below. Leakage from adjuster seal • Ensure that the adjuster seal is not damaged or twisted. If the seal is damaged, replace. • Ensure no gel or other foreign material is trapped between the seal and sealing faces on the column tube, flowcell and seal setting ring.

  • Page 33: Excessive Column Back Pressure

    • Ensure the flow tube connectors are not damaged. If so, replace. Excessive column back pressure Excessive back pressure in chromatography can result from many different areas; problems with the chromatography system, e.g. partial valve failure; the media packed bed, e.g. over compression or bed fouling;...

  • Page 34: Kits, Spare Parts And Accessories

    Chapter Kits, Spare parts and Accessories The following kits are supplied with the column: • Connection Kit • Spares Kit • Tool Kit including torque wrench It is possible to re-order these kits using the product codes listed in the table below. Connection kit A kit that contains frequently used connectors is also supplied with the column.
  • Page 35 Seal Tool - Micro S/Driver Slotted 1.2mm Spares Kit PRODUCT NUMBER DESCRIPTION SPARES Bed Support Assembly, 70 Fixed, Bed Support Assembly, 70 Adjuster, Bed Support Assembly, 100 Fixed, Bed Support Assembly, 100 Adjuster, Bed Support Assembly, 140 Fixed, Bed Support Assembly, 140 Adjuster, Adjuster seal, Evolve®70mm...

  • Page 36: Spare Parts And Kits

    Spare parts and kits The kits listed below are available to order using the product number as shown in the tables. PRODUCT NUMBER DESCRIPTION Bed Support Assembly, 70 Adjuster, SUPPORT Bed Support Assembly, 70 Fixed, Bed Support Assembly, 100 Adjuster,...

  • Page 37: Spare Parts And Kits Cont

    Pipespool Elbow 100mm Column Pipespool Elbow 140mm Column Flow Tube Seal, Ladish 3.83mm Bore Flow Tube Seal, Ladish 7.0mm Bore Bed Support Assembly, 70 Fixed, Bed Support Assembly, 70 Adjuster, Bed Support Assembly, 100 Fixed, Bed Support Assembly, 100 Adjuster,...
  • Page 38 Pipespool Elbow 100mm Column Pipespool Elbow 140mm Column Flow Tube Seal, Ladish 3.83mm Bore Flow Tube Seal, Ladish 7.0mm Bore Bed Support Assembly, 70 Fixed, Bed Support Assembly, 70 Adjuster, Bed Support Assembly, 100 Fixed, Bed Support Assembly, 100 Adjuster,...

  • Page 39: Accessories

    FLEXIBLE HOSE Flexible Hose 6mm Dia X 2m KIT(140, 200, 250) Flow Tube Seal, Ladish 7.0mm Bore AAFHK2 Click Clamp,3/4" Ladish Product DESCRIPTION Number TORQUE Torque Wrench WRENCH KIT AATWK1 (70, 100, 140) Fixed End Tool - Small (70mm,100mm,140mm Columns)

  • Page 40: Contact Us

    Chapter Contact Us To view our Evolve® Column User guides please visit: www.astreabioseparations.com/resources/evolve -column-user-guides For further enquires please contact: sales@astrea-bio.com...

  • Page 41: Appendix 1 Chromatography Performance

    Appendix Appendix 1 Chromatography Performance Performance Measurement and Column Efficiency To determine chromatographic efficiency of a packing chromatography column it is usual practice to inject a small analytical sample onto the column and monitor the response at the outlet. Typical solutions used are acetone in water or a 1M NaCl solution. If acetone solution is used then the UV absorbance is measured and plotted.

  • Page 42: Figure A1 Description Of Column Efficiency Calculations

    Figure A1 Description of Column Efficiency Calculations Asymmetry is measured at 10% of the peak height and is calculated according to the equation shown below, where a and b relate to the width of the pea section shown in Figure Asymmetry = Two further calculations can be performed to describe column efficiency.

  • Page 43: Performance Results From Evolve®Columns

    Performance Results From Evolve®Columns Performance tests have been performed on the Evolve®range of columns using Toyapearl Butyl 650M media manufactured by Tosohaas This media type has the particle size 65 micron. Accepted performance results for this media as quoted in Toyapearl Instruction Manual is expressed in terms N, number plates per m and asymmetry.

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