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Operating Instructions

Sartobind

Phenyl

nano 3 ml | 150 ml |

Jumbo 5 l

A Separation Technology
Based on Macroporous Membranes,
8 mm Bed Height

85032-540-05

Содержание Sartobind 150 ml

Страница 1: ...Operating Instructions Sartobind Phenyl nano 3 ml 150 ml Jumbo 5 l A Separation Technology Based on Macroporous Membranes 8 mm Bed Height 85032 540 05 85032 540 05...

Страница 2: ...pected before use The capsules are supplied as non sterile The membrane is dried from glycerol Intended use The products are intended and validated for single use to avoid carryover as well as tedious...

Страница 3: ...rs binding and flow properties in flow through and bind and elute purifications Sartobind 150 ml capsules have been developed for intermediate and pilot scale for the downstream processing of therapeu...

Страница 4: ...1 Venting 17 7 2 Preconditioning 19 7 3 Recommended flow rates 20 7 4 Buffer conditions 21 7 4 1 Selection of pH conditions and temperature 23 7 5 Contaminant removal from proteins in flow through mo...

Страница 5: ...50 ml and 5 l capsules 30 8 1 Installation for test 30 8 2 Operation of test 30 8 2 1 Pre washing of capsule 30 8 2 2 Diffusion measurement with Sartocheck 4 31 8 2 3 Result and evaluation 32 9 Troubl...

Страница 6: ...k place When not in use the end caps of the nano should be attached to the units to avoid oxygen all the times The sealed bag of the 150 ml and 5 l capsule should not be opened unless you use it After...

Страница 7: ...lindrical chro matographic bed and incorporated into a housing The capsules are constructed with optimized fluid channels The 150 ml and 5 l units carry a central core to minimize void volume Fluid mo...

Страница 8: ...ction chromatography HIC separates and puri fies biomolecules based on differences in their hydrophobicity The phenyl membrane adsorber follows the same rules known from the conventional hydrophobic i...

Страница 9: ...ffers with high concentrations of salt promote the adsorption of proteins on the hydrophobic membrane matrix The effect of anions and cations on protein precipitation is described in the Hofmeister se...

Страница 10: ...Proteins are eluted by decreasing the salt concentration in the elution buffer Using step or linear gradient elution proteins are eluted in the order of their hydrophobicity Regeneration and cleaning...

Страница 11: ...and 150 ml 4 bar 0 4 MPa 58 psi Jumbo 5 l 3 bar 0 3 MPa 44 psi Maximum pressure during venting 0 5 bar 0 05 MPa 7 psi Short term pH stability 2 14 Long term pH stability 3 13 see also section 5 Bindi...

Страница 12: ...Housing Polypropylene Membrane matrix Stabilized reinforced cellulose nominal pore size 3 m Membrane thickness 275 m 10 8 mils average Ligand Phenyl 12 4 Materials...

Страница 13: ...Sample protein for Sartobind Phenyl polyclonal IgG at a concentration of 1 mg ml Typical dynamic binding capacity 10 0 4 mg cm2 with 0 9 M NH4 2SO4 in 50 mM phosphate buffer pH 7 5 Unit Area Dynamic...

Страница 14: ...mm membrane layer to an internal channel and to the outlet of the capsule Install the capsule in line with a prefilter 0 2 m in front of the device Important note Capsules should be visually inspected...

Страница 15: ...up and the arrow imprinted on the banderole is pointing down We recommend to connect the Jumbo with the trolley by the three screws delivered with the trolley accessory To ensure safe unpacking the pr...

Страница 16: ...ttern inside the Jumbo 150 ml and 5 l The feed flow is from outside to inside channel 16 External channel Membrane 8 mm Central core Internal channel The direction of flow is imprinted on the banderol...

Страница 17: ...which will result in insufficient closing of the valve For appropriate venting open the vent valve screw 1 3 turn to left until all air is replaced by fluid For the 3 ml nano capsule fill a 10 20 ml L...

Страница 18: ...ed directly below the inlet do not disturb separations The capsules will function normally as long as the small air bubbles remain outside of the membrane bed Fig 4a Filling the Sartobind nano with a...

Страница 19: ...vent valves arrows 7 2 Preconditioning Prior to sample loading a sanitization and flushing procedure is recommended 1 For sanitization use 30 membrane volumes MV of 1 N NaOH solution at a flow rate of...

Страница 20: ...eline since buffers and samples have different compositions and viscosities Please test your planned flow rates using a small scale device to ensure that they fit into your pump capacities and pressur...

Страница 21: ...eraction depends mainly on salt concentrations but also on the sufficient number of exposed hydrophobic groups of the sample and on membrane ligand type and density Sample properties temperature type...

Страница 22: ...potassium phosphate pH 7 0 Choose salt concentrations as low as possible to bind the protein Higher salt concentrations may result in precipitation Commonly used salts Remarks NH4 2SO4 Typical choice...

Страница 23: ...s 7 5 Contaminant removal from proteins in flow through mode The loading conditions can be chosen to selectively retain contami nants with higher hydrophobicity and allow the target molecule with less...

Страница 24: ...nline filtering during operation With increase of pressure replace filter and restart 7 7 Washing When using capsules in bind elute mode wash with 30 ml nano 1 5 l 150 ml device or 25 l 5 l device of...

Страница 25: ...ressure to the Sartobind capsule to 2 bar The second regulator positioned immediately upstream of the Sartobind capsule should reduce the 2 bar regulated supply pressure to the 1 bar 14 5 psi draining...

Страница 26: ...merges stop the pump connect the tubing to the inlet of the Sartobind nano Make sure that no air is intro duced Remove the cap from out let Run the pump until fluid emerges from the outlet of the unit...

Страница 27: ...ication step can be scaled up to a larger capsule Recommendations Maintain Bed height automatically kept constant when using capsules with 8 mm bed height Linear flow automatically kept constant when...

Страница 28: ...resi dence time will lead to similar result Using the Sartobind nano 3 ml the scale up factor for flow rate and binding capacity is equal to a multiplication factor of membrane volumes of the listed s...

Страница 29: ...e running conditions of the Jumbo and to keep consistent upscaling increase flow rate by a factor of 1670 To assure the scale up additional experiments with the 150 ml device increase by a factor of 5...

Страница 30: ...ano 3 ml cannot be tested due to technical limitations 8 1 Installation for test Test procedure has been generated with current Sartocheck instrument family e g Sartocheck 4 plus 26288 4 16288 or 3 pl...

Страница 31: ...vent screw on top of the unit until all air is replaced by testing solvent 8 2 2 Diffusion measurement with Sartocheck 4 Choose PROGRAMMING in the main menu Choose DIFFUSION TEST Test parameters Test...

Страница 32: ...us 4 or 3 plus 8 2 3 Results and evaluation Diffusion 15 ml min Test passed diffusion value on the print out Diffusion 15 ml min Test failed red text on the print out 32 Compressed Air 3 10 bar Vent v...

Страница 33: ...do not touch the membrane bed If too much air is enclosed repeat removal as described in chapter 7 1 Venting I installed the capsule upside down Installation of capsule may be easier in the process f...

Страница 34: ...been filtered but was stored before purifica tion Proteins can form aggregates within hours or during operation Thus we recommend to prefilter inline by attaching a 0 2 m filter in front of the adsorb...

Страница 35: ...e Binding capacity is not sufficient Process optimization Use larger adsorber device or con nect two adsorbers same size in series i e connect outlet of first adsorber to inlet of second to achieve hi...

Страница 36: ...like this Technically they can be reused The reuse validation has to be performed by the user The durability of the unit depends on the nature of sample and sample preparation prefiltration as well as...

Страница 37: ...ilter before processing preferentially inline Some molecule species binds tightly and cannot be removed with 1 N NaOH 1 h Use capsule only once Proteins or contaminants are still bound from last cycle...

Страница 38: ...age overnight and longer 38 Problem Possible cause Action Change of membrane color Wrong storage No action A slight change of mem brane s color is due to oxygen and light exposure of the membrane and...

Страница 39: ...n binding capa city and flow rate Capsules and membranes are manufactured in a controlled environment The product meets all Sartorius Stedim Biotech standards for trace ability production and specific...

Страница 40: ...obind Phenyl nano 4 3 ml Luer female conectors manual 96HICP42E9BFF Sartobind Phenyl 150 ml 1 3 sanitary clamp connectors manual certificate 96HICP42E3ESS Sartobind Phenyl Jumbo 5 l 1 11 sanitary clam...

Страница 41: ...cessories Order number Description Quantity 9ZGL 0102 Trolley for Jumbo 5 l 1 stainless steel 7ZAL V0013 Reducing adapter 11 3 2 50 5 mm to 25 mm sanitary 7ZAL V0010 Reducing adapter 2 11 2 64 50 5 mm...

Страница 42: ...7 33 190 77 height diameter mm mm Frontal surface area cm2 3 6 180 Connectors Luer Sanitary 3 25 mm outer female 14 mm inner diameter Gaskets 3 internal diameter 16 mm Approximate weight 10 g 400 g Vo...

Страница 43: ...ane volume ml 5000 Dimensions 850 302 height diameter mm Frontal surface area cm2 6100 Connectors Sanitary 11 50 5 mm outer 36 mm inner diameter Gaskets 11 internal diameter 35 8 mm Approximate weight...

Страница 44: ...prior written permission of Sartorius Stedim Biotech GmbH The status of the information specifications and illustrations in this manual is indicated by the date given below Sartorius Stedim Biotech Gm...

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