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GE AxiChrom 50, Operating Instructions Manual

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GE AxiChrom 50 Operating Instructions Manual Download Page 65

Creating an Intelligent Packing
method using BioProcess media

The method wizard in UNICORN contains an Intelligent Packing option that allows the
user to create an Intelligent Packing method. Intelligent Packing methods for BioProcess
media require an initial selection of a listed medium. Thereafter the user makes appro-
priate selections to complete the method. This section describes the creation of an ex-
ample Intelligent Packing method based on a BioProcess medium. If a custom medium
is instead selected, the wizard allows a wider range of variables to be defined to optimize
the method (see Creating an Intelligent Packing method based on custom media, on
page 71).

Default values for variables in the method wizard may be changed to optimize
an Intelligent Packing method. Changes using the variables page in the Method
Editor module is for advanced users only.

Note:

The description below is for ÄKTApilot. Pages may differ slightly for ÄKTAexplorer.

1

Locate the method wizard for the relevant ÄKTA system in UNICORN.

2

On the Main page of the wizard click on the Special Method radio button, and then
select Intelligent Packing from the list. For ÄKTAexplorer, choose Intelligent Packing
in the Main Selection scrollbar.

It is recommended to click the Set Default button before a new method is
created for Intelligent Packing. This will make sure that the default packing
parameters are used.

Note:

AxiChrom 50, 70 and 100 columns Operating instructions 28-9331-08 AG

65

5 Operation

5.1 Intelligent Packing of the column with ÄKTAexplorer and ÄKTApilot

Summary of Contents for AxiChrom 50

Page 1: ...GE Healthcare Life Sciences AxiChrom 50 70 and 100 columns Operating Instructions Original instructions...

Page 2: ......

Page 3: ...5 Other equipment 36 4 6 Routine procedures 50 4 7 Over pressure situation 53 4 8 Preparing the column for a run 57 4 9 Installation in basic configuration 60 4 10 Installing the UNICORN Intelligent P...

Page 4: ...ance 149 7 2 Adapter 152 7 3 Top plate 152 7 4 Mechanical locking equipment 152 7 5 Other 153 8 Ordering information 153 8 1 Useful accessories and spare parts 155 8 2 Contact information 156 A Parts...

Page 5: ...se materials classified according to USP Class VI and FDA CFR 177 The material is free from material of animal origin or has been produced under manufacturing conditions complying with EMEA 410 01 1 2...

Page 6: ...the Appendices In some cases bold style characters either a number or letter may be contained in brackets These correspond to parts on the 100 column only that deviate from the commonly assigned item...

Page 7: ...nd NOTICES concerning the safe use of the product See definitions below Warnings WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is impo...

Page 8: ...irectives and standards that are fulfilled by AxiChrom 50 70 and 100 Manufacturing information Address of manufacturer Name of manufacturer GE Healthcare Bio Sciences AB GE Healthcare Bio Sciences AB...

Page 9: ...nd the corresponding Declaration of Conformity is valid for the instru ment when it is used as a stand alone unit or connected to other CE marked products recommended or described in the user document...

Page 10: ...ctions for installation operation and maintenance use of proper personal protective equipment and or other arrangements needed to operate the process safely For personal safety it is important that th...

Page 11: ...ing hazardous chemicals take all suitable protective measures such as wearing appropriate personal protective equipment resistant to the chemicals used Follow local regulations and instructions for sa...

Page 12: ...For details of packing and operating pressures see specifications CAUTION Use a pressure gauge pressure relief valve or other pressure safety equipment to ensure that the maximum operating pressure of...

Page 13: ...ree down to 1 m as particles can block and damage the bed supports NOTICE Handle stainless steel parts especially the bed supports with care Damage to surfaces may lead to corrosion NOTICE Chlorides a...

Page 14: ...f the columns these pressure limits shall be observed Particular attention shall be taken when connecting manual pumps to the column or deviation in normal operating procedures using KTAexplorer or KT...

Page 15: ...ted on the outer edge of the top bayonet Figure 2 1 The AxiChrom 50 70 and 100 label for serial number code number and manufacturing year Label for operating conditions The label for operating conditi...

Page 16: ...rating temperature Symbols used in safety labels Meaning Label Warning Read the user documentation before using the system Do not open any covers or replace parts unless specifically stated in the use...

Page 17: ...ced out of the slurry B The slurry forms a consoli dated bed C The adapter reaches the surface of the consolidated bed D The bed is compressed to a predetermined packing factor Axial compression is ac...

Page 18: ...ated bed height packed bed height Packing Factor PF where the consolidated bed height is read when the adapter reaches the consolidated bed where the packed bed height is read when the bed has been me...

Page 19: ...70 and 100 columns with the AxiChrom pivot stand not shown here Function Part Top mobile phase inlet outlet 1 Vent valve Hydraulic chamber hidden behind adaptor rod 2 Unpacking Hydraulic chamber outl...

Page 20: ...he hydraulic chamber inlet has an integrated check valve Bottom mobile phase inlet outlet on the bottom plate note that the column may be run upflow or downflow Port inner diameter mm of inlets and ou...

Page 21: ...ing compression there is a rise in pressure X When compression is complete UNICORN stops the flow of hy draulic liquid for some media flow conditioning will then start If selected in the Intelligent P...

Page 22: ...Operating temperature C 1 14 Operating pH range 10 20 Stainless steel bed support pore sizes m 10 20 Plastic bed support pore sizes m 16 5 19 5 10 0 11 5 6 5 7 5 Column weight empty kg 3 Short column...

Page 23: ...and 20 m 3 4 Materials The materials used to manufacture AxiChrom columns have been chosen for their bio logical and chemical compatibility with the solvents used during operation and cleaning in plac...

Page 24: ...tom plate 50 and 70 columns Yes PEEK Bed support ring Yes Stainless steel EN1 4404 316L Bed support Yes Stainless steel EN 1 4571 316Ti EN1 4462 Duplex Adapter 50 column Yes Stainless steel EN1 4404 3...

Page 25: ...PS PolyStyrene PVDF PolyVinylideneDiFluoride POM C PolyOxyMethylene PP PolyPropylene PE PolyEthylene PTFE PolyTetraFluoroEthylene For components made of Stainless steel EN1 4404 stainless steel EN 1 4...

Page 26: ...ater between the use of salt and low pH liquid This information has been collected from several published sources not from individual tests on column components It should be used only as a guide The e...

Page 27: ...2 CIP 24 h RT to 30 C 1 M pH 14 Sodium hydroxide 1310 73 2 64 17 5 CIP 3 h 1 M 20 Sodium hydroxide ethanol 7757 82 6 Adsorption 3 h 1 M4 Sodium sulfate 57 13 6 Purification CIP 5 h 8 M4 Urea Equilibra...

Page 28: ...versus other possible rinsing solutions like ethanol solutions Both the outside and the inside of the column should be thoroughly rinsed with purified water Residual chloride ions can be corrosive to...

Page 29: ...l min 35 8 ml min AxiChrom 100 The flow rate needed into the hydraulic chamber to move the adapter downwards at a given linear velocity is lower than the flow rate needed to achieve a linear velocity...

Page 30: ...rom column is delivered in two pieces Column tube assembly B with bottom plate assembly C mounted Adapter and top plate assembly A Additionally the following components are delivered AxiChrom foot wit...

Page 31: ...and outlets AxiChrom 50 column is delivered with AxiChrom foot shown in picture and AxiChrom 70 and 100 columns with the AxiChrom pivot stand not shown here Function Part Function Part Vent valve Hyd...

Page 32: ...ort Using a bed support with a larger porosity results in particles passing or getting caught in the bed support leading to poor bed efficiency and for a clogged bed support high pressure Separate 10...

Page 33: ...d support 502 Bed support top 10 m 20 m 301 Bed support bottom 10 m 20 m 302 Screw stainless steel bed support 50 70 401 End cell 100 stainless steel bed support 402 Adapter tube id 1 7 stainless stee...

Page 34: ...r in the exploded diagrams in Appendices A C 4 4 Wetting instructions for plastic bed supports Follow these instructions to wet top and bottom plastic bed supports before packing the column 1 Before u...

Page 35: ...round the cir cumference 7 If necessary place the adapter with the bed support downwards into a container of 20 ethanol to prevent air from entering the flow path 8 Mount the adapter on to the column...

Page 36: ...ications on page 22 4 6 Routine procedures The following sections describe procedures that are routinely performed when operating the column and or servicing the column These sections are cross refere...

Page 37: ...nd of the stand arm Fitting the column onto the AxiChrom pivot stand Assemble the column onto the pivot stand as follows 1 Make sure that the pivot stand is placed on an even foundation 2 Make sure th...

Page 38: ...wn position to facilitate removal of the bottom plate WARNING Keep fingers away from the locking mechanism while rotating the column to avoid injury Tilting the column tube 1 Release the pin at the ba...

Page 39: ...information The locking equipment allows the position of the adapter to be locked without the need for hydraulic pressure CAUTION High pressure Never move the adapter in a column while mechan ical loc...

Page 40: ...to compress the bed further than what was intended during the packing of the column 5 Check that the mechanical locking equipment is firmly fixed onto the column and that the parts sit securely 6 To...

Page 41: ...t the top mobile phase inlet outlet 3 Remove the nuts and withdraw the mechanical locking equipment from the column Fitting the top plate and adapter The following procedure is used to fit the top pla...

Page 42: ...out 10 cm from the adapter Grip the adapter rod with one hand on each side of the top plate 5 Carefully insert the adapter into the mouth of the column tube at an angle so that the adapter enters into...

Page 43: ...between the top plate and the adapter or top plate and top bayonet when the top plate slides down the adapter rod CAUTION To avoid use of excessive force and risk of trapping fingers lu bricate the a...

Page 44: ...ar head of liquid 1 2 cm above the top surface of the settling slurry before inserting the adapter into the column tube NOTICE Never force the adapter into the column manually Only use hydraulic press...

Page 45: ...aligned with the machined mark at the left close open Figure 4 5 Opening and closing of the bayonet WARNING Ensure that the top plate is completely closed by confirming that the tool in the top plate...

Page 46: ...outlet in the bottom plate and connect a tubing from this outlet to waste 2 Ensure that the inlets outlets in the top plate including the top mobile phase in the adapter rod are sealed 3 Connect one e...

Page 47: ...op the pump This is to facilitate safe removal of the liquid above the adapter 6 Ensure that the hydraulic chamber outlet and or the hydraulic chamber vent valve are opened 7 Ensure that the column is...

Page 48: ...the adapter rod Limitation of adapter movement in the long column tube The length of the adapter rod in a long column tube is designed for packing bed heights from 30 cm to 50 cm The adapter rod leng...

Page 49: ...is stable in the mouth of the column or hold the adapter in an upright position 6 Dampen the adapter rod 802 using a cloth wetted with 20 v v ethanol and carefully slide the top plate approximately 15...

Page 50: ...ntelligent Packing and UNICORN at KTApilot is not verified as a safety monitoring system for the columns Causing over pressure An over pressure situation can occur due to Situation Error type Operator...

Page 51: ...dure 1 Stop the pump 2 Alert people in the room that the column is over pressurized 3 Monitor the pressure 4 Take all necessary safety precautions before depressurizing the column 5 Try to release the...

Page 52: ...lic chamber vent valve 107 and hold it in place 2 Open the hydraulic chamber vent valve and collect the liquid in the syringe until the excess pressure in the hydraulic chamber is normalized Do not to...

Page 53: ...bing and plug the top and bottom inlets outlets and the hydraulic chamber inlet before the column is moved NOTICE Do not hold the column by the adapter rod as this can damage the column NOTICE Take ca...

Page 54: ...e mechanical locking equipment when moving a packed column Note CAUTION Make sure not to touch the locking equipment for the arm of the pivot stand when moving the column to avoid injury and rotation...

Page 55: ...the X direction i e the tilt of the whole back section between the wheels The other two points are the adjustable feet b and c at the front of the stand and allow levelling in the Y direction 1 Screw...

Page 56: ...ample water and or 20 v v ethanol to remove any surface residues acquired during removal from the packaging Before use carefully pump 1 0 M NaOH into the column empty and then pump a washing liquid fo...

Page 57: ...ing kits and connections on page 186 Installation to KTAexplorer For details about tubing see section Tubing used between column and system on page60 Column Valve V2 Column Valve V3 2 3 2 3 Figure 4 8...

Page 58: ...A avant 150 Connect the column to the system as described in the illustration in the Intelligent Packing phase and according to those column valves chosen in the method see step 6 in section Creating...

Page 59: ...9888 89 Tubing kit AxiChrom 50 KTA avant desk Pos id 1 7 L 1600 x2 id 1 7 L 1200 x2 1 id 1 7 L 1200 x1 id 1 7 L 800 x1 2 id 2 9 L 1600 x1 id 2 9 L 1600 x1 3 id 1 7 L 200 x2 id 1 7 L 200 x2 4 AxiChrom...

Page 60: ...ntelligent Packing strategy on KTAexplorer and KTApilot Install the correct UNICORN strategy containing Intelligent Packing for the appropriate system according to the instructions on the cover of the...

Page 61: ...KTA explorer 1 Open the UNICORN manager module 2 Under Administration select System Setup 3 Select the appropriate system and click on Edit System 4 Select the appropriate strategy and then click on C...

Page 62: ...ake it possible to detect leakage at an early stage with minimized consequences and the risk for potential leakage of hazardous liquids is diminished Note Before packing the column pump 1 0 M NaOH thr...

Page 63: ...ed These require that flow occurs into the process chamber via the mobile phase inlet outlet By contrast during packing the method is con structed so that the air trap is bypassed as hydraulic liquid...

Page 64: ...repack the column or c Filter smaller volumes of medium using a vacuum filtration system with a glass filter funnel Slurry volume The method wizard calculates and presents the needed slurry volume in...

Page 65: ...ault values for variables in the method wizard may be changed to optimize an Intelligent Packing method Changes using the variables page in the Method Editor module is for advanced users only Note The...

Page 66: ...propriate material The only available option is Glass Column material Select the appropriate bed support type and material The mate rial options are stainless steel and plastic The porosity choice 10...

Page 67: ...ed range Note There are different scales for stainless steel and plastic bed supports on the column Target bed height Select the appropriate inlet Inlet for hydraulic chamber liquid Select the appropr...

Page 68: ...cking Factor Unchecking this option for example for custom media see Creating an Intelligent Packing method based on custom media on page 71 allows the user to enter a Packing Factor in the range of 1...

Page 69: ...and test liquid for the selected medium Equilibration volume Leave checked to use the suggested default liquid velocity of 30 cm h Use default liquid velocity during equilibration Leave checked to use...

Page 70: ...f flow condition ing 6 Click on Next to display the Column Packing Summary wizard page Figure 5 5 Intelligent Packing Wizard 5 of 5 KTApilot This page provides a summary of the options selected in the...

Page 71: ...Packing methods may also be developed for custom media using the wizard The wizard facilitates this by providing an extended range of variables that may be modified to optimize the method For example...

Page 72: ...zard NOTICE Over compression during packing can damage the medium In the Intelligent Packing tests performed by GE Healthcare on its base media selection the default Packing Factor for each medium has...

Page 73: ...eral times Gently tap the column and shake the tubing while flowing packing liquid into the column to help free the air bubbles Drain the column through the bottom phase inlet until 1 cm of liquid rem...

Page 74: ...on of the media stirrer and additional liquid volume from rinsing Note 5 Add more packing liquid into the column right up to the edge of the glass column tube Medium stuck on the column wall or in the...

Page 75: ...not turn over by accident CAUTION Make sure that the top plate is completely closed by confirming that the bayonet tool in the top plate is aligned with the ma chined mark on the top bayonet 2 Make su...

Page 76: ...ion NOTICE Ensure that the column is free from air before the process run is started NOTICE Do not seal the hydraulic chamber vent valve too early i e wait for the instruction to close the vent valve...

Page 77: ...k on Continue in the wizard when the adapter comes into contact with the consolidated bed surface to achieve the right compression Clicking continue too early or too late will deliver a too low or too...

Page 78: ...e different scales for stainless steel and plastic bed supports on the column Note 5 When the packing test is completed open the Evaluation module of UNICORN and perform an integration of the peak usi...

Page 79: ...nsure that the air trap is filled with mobile phase liquid This is because the methods created using the Intelligent Packing wizard are constructed so that the airtrap is in line with the column when...

Page 80: ...For more information on Column Handling refer to the documentation for your UNICORN software Open the Column Handling dialog To open the Column Handling dialog select Tools Column Handling in any of...

Page 81: ...e Parameters tab in the Column Handling dialog click 1 Result The New Column Type dialog opens Note The Intelligent Packing method is only compatible with AxiChrom columns AxiChrom 50 70 and 100 colum...

Page 82: ...how medium types by technique drop down list above Result The following parameters are automatically filled in can be edited if appropriate 2 Enter the remaining parameter values for the new column ty...

Page 83: ...database and displayed in the Column types list Note If traceability of result is requested a column individual can be created under column type for logging of the data see UNICORN 6 1 Method Manual...

Page 84: ...ent Packing method using GE approved media This section describes the creation of an example Intelligent Packing method based on GE approved media If custom media is selected instead the method settin...

Page 85: ...layed in the Method Outline pane to the left and the default settings for each of the phases are shown in the Phase Properties pane to the right AxiChrom 50 70 and 100 columns Operating instructions 2...

Page 86: ...llustration below shows the Method Outline pane to the left and the Phase Properties pane of the Method Settings phase to the right Make sure that all parameter values are correct Note If the check bo...

Page 87: ...Intelligent Packing phase 4 The illustration below shows the Method Outline pane to the left and the Phase Properties pane of the Intelligent Packing phase to the right Note If the user wants to make...

Page 88: ...d packing buffert for the selected medium is also shown in the dialog 5 Result The Suggested Slurry Recipe dialog opens Note The Slurry Recipe button is not available in Custom Packing settings Click...

Page 89: ...e How to connect the column to the system dialog opens Note The text will be updated dependent on the column positions chosen but the illustration will remain the same Click OK to close the dialog Axi...

Page 90: ...olumn volumes 7 The illustration below shows the Method Outline pane to the left and the Phase Properties pane of the Equilibration phase to the right Note All parameters in this phase can be changed...

Page 91: ...quality The eluate is monitored by measuring the UV absorption at 280 nm or the conductivity and the curve can be evaluated in Evaluation See Performance eveluation of the column in ManualNameComplete...

Page 92: ...ay be modified to optimize the method For example it is possible to Pack according to a desired Packing Factor or bed height If selecting the latter option the user stops the compression when the fina...

Page 93: ...TAavant Follow the instructions in this Operating Instructions to prepare the slurry prime the column pour slurry into the column and to pack the column The UNICORN pages may differ slightly for KTAav...

Page 94: ...uctions see below CAUTION Ensure that the pivot stand locking pin is secured before starting the process so that the column cannot turn over by accident 1 Prepare the media slurry see Preparing the sl...

Page 95: ...e adapter into the mouth of the column tube at an angle to ensure that no air is trapped in the column 7 Place a syringe without plunger onto the hydraulic chamber vent valve and open the vent valve 8...

Page 96: ...ng down and the media is packed NOTICE Make sure to remove the syringe otherwise it may become hooked under the adapter rod top and hinder the packing es pecially if packing a low bed height 11 Note w...

Page 97: ...utes The instruction is for KTApilot The commands differ slightly for KTAex plorer Note 15 Run the column upflow for a few minutes to remove residual air possibly trapped in the adapter The column is...

Page 98: ...king factor 2 Prime the column see Priming the column on page73 and close the bottom mobile phase inlet outlet 3 Pour the slurry into the column see Pouring slurry into the column on page 74 4 Fit the...

Page 99: ...ifferent scales for stainless steel and plastic bed supports on the column Note 11 Continue to compress the bed until the target bed height is reached Calculate the achieved Packing Factor PF or Conti...

Page 100: ...the packing is evaluated Note The sample volume should be approximately 1 to 2 of the total bed volume and 1 to 2 v v acetone can be used Alternatively 0 8 M sodium chloride or equivalent can be appl...

Page 101: ...acceptable A change in the shape of the peak is usually the first indication of bed deterioration due to use The asymmetry factor As describes the deviation from an ideal Gaussian peak shape and is c...

Page 102: ...uid has accumulated under the packed bed increase the flow rate successively again monitoring the pressure carefully During this procedure the adapter will move upwards in the column and the medium be...

Page 103: ...and the expected working life cycle of the medium and equipment For more information about which chemicals to use for CIP see the table on page 27 Ensure that the media and the column withstands the c...

Page 104: ...m h b 2 column volumes downflow at a velocity of 60 cm h 3 For 50 and 70 columns with plastic bed support Recirculate 5 column volumes of 1 0 M NaOH upflow for 2 hours followed by downflow for 2 hours...

Page 105: ...column on page 102 2 Remove the adapter and top plate assembly see Removing the top plate and adapter on page 49 3 Release the pin at the pivot stand if used tilt the column and pour out medium into a...

Page 106: ...fit the mechanical locking equipment see Mechanical locking on page 39 onto the column Plug the mobile phase inlets outlets and the hydraulic chamber inlet Ensure that the storage conditions of the co...

Page 107: ...e 803 placed at the end of the adapter rod right below the adapter rod top 105 Ensure that the top plate rests on its planar side to keep it in place For the stainless steel bed support ensure that th...

Page 108: ...valve performing a drainage test performing a column leakage test Referencingcolumncomponents Many of the procedures in this Operating Instructions make reference to specific com ponents on the AxiChr...

Page 109: ...been emptied and properly cleaned If this is not pos sible wear protective clothing CAUTION When handling the adapter and top plate assembly ensure that fingers or hands are not caught in between the...

Page 110: ...5 2 5 3 4 Allen key mm 13 10 Torque wrench key with socket mm 2 5 Torque driver with bit mm Yes x2 Bayonet tool1 Yes x2 Centering tool1 Yes Gloves Yes for lubrication of friction surfaces Bottle of 20...

Page 111: ...s are replaced once a year The scraper seal on the adapter bed support shall be regularly checked for scratches and other damage see Servicing the stainless steel adapter bed support on page112or Serv...

Page 112: ...the bed support with water and then inspect for signs of damage If necessary replace the entire bed support 6 Clean the bed support if required for example due to clogging as determined by a drainage...

Page 113: ...d water 5 Test the cleaning efficiency with a drainage test Reassembling the stainless steel adapter bed support 1 Fit the O ring 603 411 into the groove in the adapter 501 for the 50 and 70 columns o...

Page 114: ...out from the bed support wall and moving it downwards a fraction and then repeating on an adjacent section of the O ring and so on The scraper seal can be mounted in a plastic container filled with w...

Page 115: ...the mounting tool on top of the O ring when in place in the groove and then draw the mounting tool around the circumferance of the O ring while applying pressure downwards Draw the mounting tool in b...

Page 116: ...he 100 columns Tightening of the screws 401 will cause the O ring 603 411 to be pushed onto the bed support and damage the bed sup port Approx 1 mm A B 4 For the 100 column only Check and replace the...

Page 117: ...m their holes b There are three additional threaded holes on the bottom plate assembly Use three of the removed bolts 801 and insert them into these three holes c Gently screw in each bolt a little at...

Page 118: ...3 Refit the bed support 302 onto the bottom plate 502 being careful to align the holes around the circumference on both parts NOTICE The bed support must be mounted in the position according to figur...

Page 119: ...removed from the column see Removing the top plate and adapter on page49 It is also required that the adapter bed support is removed for this operation see Servicing the stainless steel adapter bed su...

Page 120: ...ge to unscrew the adapter from the adapter rod 3 Gently pull on the adapter to withdraw the tubing from inside the adapter rod Figure 6 7 Withdrawing the tubing from inside the adapter rod 120 AxiChro...

Page 121: ...nd if nec essary replace the O rings 127 and 411a Figure 6 9 Unscrewing the top end cell on AxiChrom 100 column Allowing the O ring 411a to remain on the end cell facilitates removal Note 6 Slide the...

Page 122: ...O ring and roll it onto the adapter d Remove the Parafilm Reassembling the adapter rod and top plate 1 For the 100 column only Screw the top end cell 402 back by hand Ensure that it is screwed tight...

Page 123: ...6 If necessary remove and replace the O ring 104 in the adapter rod top This O ring does not normally suffer from wear and its replacement is a difficult procedure Note 7 Fit the adapter rod top onto...

Page 124: ...the support net 422a as well B A Figure 6 11 Remove the snap ring and the bed support from the distributor by hand A or using the removal tool B To service or change distributor scraper seal and adapt...

Page 125: ...y 6 Remove the scraper seal 605 and O ring 604 by gently moving them up the dis tributor a bit at a time around the circumference The O ring is secured within the scraper seal so they will most likely...

Page 126: ...he flat surface face downwards until it sits on the distributor flange B c Fit the O ring 604 by gently moving it down the distributor a bit at a time around the circumference Use 20 v v ethanol for l...

Page 127: ...ve created by the scraper seal and the wall of the distributor Press down one part of the O ring and then at an opposite point on the O ring circumference Continue until the O ring is located within t...

Page 128: ...d that has the connector with a smaller diameter shall be connected to the system end see exploded diagrams in Appendix A C according to the column type Note 4 Feed the free end of the adapter tube th...

Page 129: ...sulting from the adapter tubing coming loose Figure 6 18 Tightening the stop screws 10 a For 50 and 70 columns Flush the adapter downflow with 20 ethanol to ensure that the adapter and tubing are fill...

Page 130: ...it the bed support and snap ring assembly onto the distributor Ensure that the bed support is in contact with the distributor all around the circumference Check that the snap ring is correctly and eve...

Page 131: ...les on the bottom plate assembly Use three of the removed bolts 801 and insert them into these three holes c Gently screw in each bolt a little at a time to jack the bottom plate assembly C until it s...

Page 132: ...he distributor all around the circumference Check that the snap ring is correctly and evenly mounted around the whole circumference of the distributor Note Figure 6 22 Fitting the bed support in the s...

Page 133: ...at a time crosswise Do not overtighten 5 Lubricate the inside of the tube end 201 and the O ring 606 of the bottom plate assembly C and refit it onto the column Tighten the screws crosswise and a lit...

Page 134: ...everage to unscrew the adapter from the adapter rod 2 Slide the top plate 803 down the length of the adapter rod 802 and remove Apply a small amount of ethanol 20 v v on the adapter rod for easier mov...

Page 135: ...ol 20 v v on the adapter rod for easier movement of the top plate along the adapter rod NOTICE Always wear gloves when handling the adapter rod to avoid fat from unprotected fingers and scratching the...

Page 136: ...emove the nuts 806 and washers 119 connected to the tie bars underneath the bottom flange 805 Carefully remove the bottom flange 4 Unscrew 1 2 tie bars see Replacing tie bars on page 139 5 Carefully r...

Page 137: ...anar surface on the outer rim of the bottom flange is aligned with the planar surface of the top bayonet and that the holes for the pivot stand are aligned 4 Place the washers 119 onto the tie bars at...

Page 138: ...ol into the top A and bottom B of the column It is important that the centering tools are placed into the column tube to the correct marking point at the respective ends otherwise the column will not...

Page 139: ...oosen and unscrew the damaged tie bar s 901 connected to the top bayonet 804 2 Screw in a new tie bar to one of the tie bar holes by hand and tighten using a wrench at the key handles One of the ends...

Page 140: ...ve represents the flow di rection and shall be fitted so that it points inwards to the column Figure 6 27 Positioning of the check valve on AxiChrom 50 or 70 column CAUTION To avoid risks caused by ov...

Page 141: ...mn Figure 6 28 Positioning of the check valve on AxiChrom 100 column CAUTION To avoid risks caused by over pressure the check valve shall be fitted into the valve housing such that the direction of th...

Page 142: ...harm the bed support Use a new bed support as reference and the same weight as used with the used bed support Note 5 Open the bottom outlet and record the time it takes for the liquid level to move ev...

Page 143: ...or the hydraulic chamber Other wise the correct pressure is not registered during measurement by the check valve at the hydraulic chamber inlet Note 7 Close the hydraulic chamber vent valve when only...

Page 144: ...ble height Water should emerge from the open mobile phase inlet outlet 7 Assemble the mechanical locking equipment see Mechanical locking on page 39 8 Pump liquid into the process chamber at a low liq...

Page 145: ...uble peaks The extra peak is before the main peak i e leading Try to compress the bed slightly and retest Otherwise repack the column with a higher Packing Factor Cracks in the bed or a gap between th...

Page 146: ...luation Decreased leading Repack the column with a higher or lower Packing Factor Bed packed too densely or too loosely Poor stability when repeating packing evaluation Decreasing number of plates Ens...

Page 147: ...e and replace if necessary Leaking bottom bed support Liquid velocity should be in the range 15 60 cm h depending on particle size The smaller the particle the higher the velocity Liquid velocity too...

Page 148: ...se buffer before starting the Intelligent Packing method A dip in the baseline during performance evaluation Ensure that no connections are leaking Change gaskets if necessary Leakage in the tubing be...

Page 149: ...instructions underServicing the adapter rod adapter tubing and top plate stainless steel bed support on page 119 Adapter does not move upwards in the column despite applied pressure upflow in the proc...

Page 150: ...the Intelligent Packing wizard and be careful to add the right volume of slurry Wrong amount of medium added to the column Check that defaults values were used during Intelligent Packing Performance o...

Page 151: ...on page 119 The parts can loosen after using the column several times Check that the adapter rod is tight before performing each packing procedure The adapter is not tight enough The adapter is parti...

Page 152: ...adapter down by 1 2 mm Loosen the nuts Make sure not to over pressurize the column or the bed Nuts at the mechanical locking are too tight to loosen by hand 7 5 Other Action Comment Problem Replace da...

Page 153: ...28 4018 41 EN 1 4401 1 Mechanical locking 100 Yes 28 9231 80 PP 1 Media stirrer 40 Yes 28 9191 03 PP 1 Media stirrer USP 6 80 mm Yes 28 9055 41 PVDF 1 Tubing kit AxiChrom 50 KTAexplorer desk Yes 28 90...

Page 154: ...Stop plug 5 16 No 18 1127 76 PEEK 3 Union 5 16 female M6 male Yes 28 9608 75 PEEK 1 Vent valve M8 No 19 5170 01 PP PTFE 2 Stop plug M6 male No 18 1142 08 8 Union 5 16 female 1 16 male No 19 2145 01 ET...

Page 155: ...ust be ex changed The bed support kits are numbered 28 3664 46 through 28 3664 49 Note Spare parts recommended to keep on site It is recommended to keep the following spare parts on site Then keep the...

Page 156: ...08 Tube connection 5 16 id 1 7 Fem fem 109 O ring 11 1x1 6 111 O ring 54x2 114 a b Tube support 50 115 a b Check Valve holder 116 O ring 3 6x1 6 117 a b Check Valve 118 Washer 6 4x12 119 AxiChrom foot...

Page 157: ...8 3x1 78 410 O ring 34 646x1 78 411 Washer 3 2x9x0 8 412 O ring 2 5x1 3 413 O ring 2 5x1 3 416 Snap ring 50 421 a b Support net 422 a b Adapter 50 501 Bottom plate 50 502 O ring 3x1 602 a b O ring 44...

Page 158: ...D Adapter assembly E Bayonet tool Hook wrench key Mounting aid for o rings Centering tool 50 Diagrams Meaning Symbol Number corresponds to Pos no in Material conformity and Spare part list No Illustra...

Page 159: ...Figure A 1 Column and foot AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 159 A Parts list and diagrams AxiChrom 50 column...

Page 160: ...Figure A 2 Column tube adapter and bottom plate 160 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG A Parts list and diagrams AxiChrom 50 column...

Page 161: ...Figure A 3 Adapter assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 161 A Parts list and diagrams AxiChrom 50 column...

Page 162: ...Figure A 4 Adapter assembly plastic bed support 162 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG A Parts list and diagrams AxiChrom 50 column...

Page 163: ...gure A 5 Adapter plate assembly and bottom plate assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 163 A Parts list and diagrams AxiChrom 50 colu...

Page 164: ...Figure A 6 Adapter plate assembly and bottom plate assembly plastic bed support 164 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG A Parts list and diagrams AxiChrom 50 column...

Page 165: ...Figure A 7 Column tube assembly AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 165 A Parts list and diagrams AxiChrom 50 column...

Page 166: ...ring 65x3 108 Tube connection 5 16 id 2 9 Fem fem 109 O ring 14X1 5 111 O ring 72x2 114 a b Tube support 70 115 a b Check Valve holder 116 O ring 3 6x1 6 117 a b Check Valve 118 Washer 6 4x12 119 O ri...

Page 167: ...5x1 3 413 Snap ring 70 421 a b Support net 422 a b Adapter 70 501 Bottom plate 70 502 O ring 3x1 602 a b O ring 63 2x2 8 EPDM 604 Scraper 70 605 O ring 63 09x3 53 606 Pivot stand 50 70 100 300 701 1...

Page 168: ...r assembly E Bayonet tool Hook wrench key Mounting aid for o rings Centering tool 70 Diagrams Meaning Symbol Number corresponds to Pos no in Material conformity and Spare part list No Illustrating par...

Page 169: ...Figure B 1 Column and pivot stand AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 169 B Parts list and diagrams AxiChrom 70 column...

Page 170: ...Figure B 2 Column tube adapter and bottom plate 170 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG B Parts list and diagrams AxiChrom 70 column...

Page 171: ...Figure B 3 Adapter assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 171 B Parts list and diagrams AxiChrom 70 column...

Page 172: ...Figure B 4 Adapter assembly plastic bed support 172 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG B Parts list and diagrams AxiChrom 70 column...

Page 173: ...gure B 5 Adapter plate assembly and bottom plate assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 173 B Parts list and diagrams AxiChrom 70 colu...

Page 174: ...Figure B 6 Adapter plate assembly and bottom plate assembly plastic bed support 174 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG B Parts list and diagrams AxiChrom 70 column...

Page 175: ...Figure B 7 Column tube assembly AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 175 B Parts list and diagrams AxiChrom 70 column...

Page 176: ...ge valve polished 107 O ring 98x3 108 Tube connection 5 16 id 2 9 Fem fem 109 O ring 23 52x1 78 111 O ring 104 5x3 114 a b Tube support 100 115 a b Check valve holder 116 Check valve single cpl 118 Wa...

Page 177: ...be id 2 9 plastic bed support 405 1 2 3 O ring 70x2 5 411 a b Washer 4 3x12 1 SS 412 O ring 3 69x1 78 413 Snap ring 100 plastic bed support 421 a b Adapter plate 100 SS bed support 501 Adapter plate 1...

Page 178: ...ttom plate assembly C Adapter assembly with tube D Adapter assembly E Bayonet tool Centering tool 100 Mounting aid for o rings Hook wrench key Diagrams Meaning Symbol Number corresponds to Pos no in M...

Page 179: ...701 4 701 3 701 1 701 2 131b 131a Figure C 1 Column and pivot stand AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 179 C Parts lists and diagrams AxiChrom 100 column...

Page 180: ...A 801 B C Figure C 2 Column tube adapter and bottom plate 180 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG C Parts lists and diagrams AxiChrom 100 column...

Page 181: ...106 107 103 101a 602a 602b 102 405 1 405 3 405 2 5x D E Figure C 3 Adapter assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 181 C Parts lists a...

Page 182: ...105 106 107 103 101a 602a 602b 102 405 3 405 2 5x 405 1 D E Figure C 4 Adapter assembly plastic bed support 182 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG C Parts lists and d...

Page 183: ...412 413 127 402 411a 411b 404 Figure C 5 Adapter plate assembly and bottom plate assembly stainless steel bed support AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 183 C Parts li...

Page 184: ...3 412 413 402 404 606 421a 421b 127 Figure C 6 Adapter plate assembly and bottom plate assembly plastic bed support 184 AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG C Parts list...

Page 185: ...1 101b 116 118 124 115a 114a 804 115b 114b 805 119 806 Figure C 7 Column tube assembly AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 185 C Parts lists and diagrams AxiChrom 100 c...

Page 186: ...00 id1 2 L 800 2 Top valve P2 KTAexplorer Column hydraulic chamber inlet id1 2 L 1100 id1 2 L 1100 id1 2 L 500 3 Column hydraulic chamber outlet waste id2 9 L 1600 id2 9 L 1600 id2 9 L 1600 id2 9 L 16...

Page 187: ...3 6 7 1 5 4 3x 8 7 2 9 8 9 9 7 1 3 7 9 2 4 9 7 9 10 10 Figure D 1 Kits and connections for KTAexplorer AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 187 D Tubing kits and connect...

Page 188: ...L 1600 id1 7 L 1600 id1 7 L 1600 3 For use with Valve 5 16 straight id2 9 L 200 id1 7 L 200 id1 7 L 200 4 5 16 female both valve ends 28 9076 74 Valve 5 16 straight 53 To be mounted at Pos 4 tubing en...

Page 189: ...4 5 1 2 1 2 1 3 1 2 3 1 6 Figure D 2 Kits and connections for KTApilot AxiChrom 50 70 and 100 columns Operating instructions 28 9331 08 AG 189 D Tubing kits and connections...

Page 190: ...ing 117 C Check valve servicing 140 141 CIP 103 Cleaning 56 Adapter bed support 112 Bottom bed support 117 CIP 103 Column Accessories 153 KTAexplorer connec tion 57 KTApilot connection 58 AxiChrom piv...

Page 191: ...pressure 22 O ring Adapter servicing 113 126 O ring replacement 112 Over pressure 51 P Packing Factor Wizard setting 68 Packing list 30 Packing pressure 22 Packing the column Intelligent Packing 75 Ma...

Page 192: ...let 31 Top plate Fitting 41 Removing 49 Top plate servicing 119 124 Tubing kits 186 U UNICORN Strategy installation 60 Unpacking a packed bed 102 W Wizard Slurry concentration 69 Wizard for Intelligen...

Page 193: ......

Page 194: ...ormation UNICORN Any use of this software is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products A copy of this Standard Software End User License...

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