Related Manuals for Sartorius Stedim Biotech Sartobind Q Series
Summary of Contents for Sartorius Stedim Biotech Sartobind Q Series
- Page 1 Operating Instructions ® Sartobind Q and S Membrane Adsorbers Void Volume Optimized Capsules With 4 | 8 mm Bed Height nano 3 mL 75 mL | 150 mL 1.2 L Jumbo 5 L 85037-545-36...
- Page 2 Read operational instructions carefully before using Sartobind capsules. Use of the products in applications not specified or not described in this manual, may result in improper function, personal injury, or damage of the product or material. The capsules are supplied as non-sterile.
- Page 3 La utilización de este producto en aplicaciones ajenas o no establecidas en el manual de operación, puede provocar un mal funcionamiento del pro- ducto, del material, así como daños personales. Las cápsulas suministradas en este producto no son estériles. La membrana es de secado de Glicerina. L’utilizzo dei prodotti per applicazioni non specifi- cate o non descritte in questo manuale, può...
- Page 4 把产品用于手册中没指定或描述的应用， 可能导 致产品失效，人员受伤，或使产品及物料受损。 囊式膜柱以非灭菌的形式供应。膜用甘油脱水。 当製品を該当しない用途、 あるいは当製品取扱説明 書に記載されていない応用分野において使用した 場合、 当製品の機能上の不具合や損傷、 人体への危 害、 あるいは他の物品の損傷を招く恐れがあります。 当カプセル製品は滅菌処理されていません。 当メン ブレンはグリセリンを用いて乾燥させてあります。 Intended use The products are intended and validated for single use to avoid carryover as well as tedious and costly cleaning validation procedure. However it is technically possible to reuse after cleaning in place depending on application, character of sample and process.
- Page 5 Sartobind nano 3 mL capsules have been developed for working with small sample volumes while retaining the cylindrical design of large scale Membrane Adsorber. They are perfect for small scale applications, and also for screening purposes and laboratory-scale bind and elute and flow through purifications.
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Page 6: Table Of Contents
Table of Contents 1. Storage conditions ..............8 2. Introduction ................8 3. Technical data ............... 10 4. Materials ................12 5. Binding capacity ..............12 6. Installation ................13 7. Operation ................15 Venting ................. 15 Cleaning................ 16 Autoclaving ..............17 Recommended flow rates .......... - Page 7 8. Testing of membrane samples enclosed with 5 L capsules ..............25 Testing Sartobind Q 30 mm discs ....... 25 8.1.1 Materials for test of Q membrane discs ... 25 8.1.2 Method for test of Q membrane discs ..... 26 8.1.3 Calculation of membrane binding capacity ..
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Page 8: Storage Conditions
1. Storage conditions Sartobind capsules should be stored clean, dry and away from direct sunlight in the box at room temperature. Store membrane samples enclosed in Jumbo shipment in a safe place as they cannot be ordered again. 2. Introduction The capsules with 4 and 8 mm bed height are ion exchange chromatography devices based on macroporous membranes. - Page 9 costly cleaning validation procedures. However, it is technically possible to reuse after cleaning in place (see also section 7.11). Their major application is for single use as they are validated for contaminant removal from proteins in flow-through mode (negative chromatography) to bind DNA, residual protein, host cell proteins, endotoxins, viruses and aggregates.
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Page 10: Technical Data
3. Technical data Membrane volume (MV) 3 mL Nominal membrane area 110 cm Bed height 8 mm Design Cylindrical Sartobind Q typical 10% dynamic binding capacity* 88 mg Sartobind S typical 10% dynamic binding capacity* 77 mg Maximum pressure bar (Mpa, psig) at 20°C 4 (0.4, 58) Maximum pressure during venting –... - Page 11 75 mL 150 mL 1.2 L 2,700 cm 5,500 cm 44,000 cm 182,000 cm 4 mm 8 mm 8 mm 8 mm Cylindrical Cylindrical Cylindrical Cylindrical 2.16 g 4.4 g 35 g 145 g 1.89 g 3.9 g 31 g 127 g 4 (0.4, 58) 4 (0.4, 58)
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Page 12: Materials
4. Materials Housing Polypropylene Membrane matrix Stabilized reinforced cellulose nominal pore size >3 µm Ion exchange ligand Strong basic anion exchanger: Sartobind Q quaternary ammonium (R-CH Ion exchange ligand Strong acidic cation exchanger: Sartobind S sulfonic acid (R-CH 5. Binding capacity Data are based on dynamic binding capacity measurements 10% using 3 layers of 5 cm membrane discs (15 cm... -
Page 13: Installation
6. Installation The contents of the package is described in chapter 12. When unpacking capsule, protect inlet and outlet connectors from damage. Never keep or place the capsule directly on the floor on the connectors. This might damage the sanitary adapters. For unpacking of Jumbo 5 L capsule, take the capsule including the styrene foam end protectors, out of the box and place it upright on the end protectors. - Page 14 Capsules should be visually inspected before use. In case of visible damage, the capsule should be replaced. Close vent valves before use by screwing the valve clockwise. The direction of flow is imprinted on the banderole of the unit. External channel Membrane (4 | 8 mm) Central core Internal channel...
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Page 15: Operation
7. Operation 7.1 Venting It is important to remove air from the unit completely. The 75 mL, 150 mL, 1.2 L and 5 L capsules carry vent valves. The vent valves are equipped with hose barb connectors for the fluid spilled out during venting. -
Page 16: Cleaning
Very small air bubbles observed directly below the inlet of the nano do not disturb separations. The capsule will function normally as long as the small air bubbles remain outside of the membrane bed. Outlet Inlet Fig. 6: Filling the Sartobind nano with a Luer syringe 7.2 Cleaning The modules have to be cleaned in place directly after unpacking... -
Page 17: Autoclaving
7.3 Autoclaving The capsules can be autoclaved once at 121°C for 30 minutes at 1 bar (0.1 MPa| 14.5 psi). Prewet the capsule with equilibration buffer. Do not use pure water. Close valves immediately after sterilisation. For auto- claving Sartobind Jumbo refer to separate autoclaving instruc- tions enclosed in delivery. -
Page 18: Buffer Conditions
7.5 Buffer conditions In the majority of applications an equilibration buffer concentra- tion of 10 mM provides sufficient buffering capacity and prevents the protein of interest from precipitation. The ionic strength should be kept as low as possible to avoid reduction of binding capacity. -
Page 19: Selection Of Ph Conditions
7.6 Selection of pH conditions In ion exchange chromatography a charged molecule is bound to oppositely charged groups attached to the insoluble matrix. This binding is reversible by application of salt ions to the buffer eluting the molecule. The pH value at which a biomolecule has no net charge is the isoelectric point: pI. -
Page 20: Sample Preparation
To remove contaminating proteins and aggregates with Sartobind S in flow through mode, process impurities have to be charged positively to bind to S while the target protein stays negative as the pH of the buffer is above the pI of the protein product and flows through without binding. -
Page 21: Elution
7.10 Elution To elute the target protein use buffers with appropriate salt concentration. 7.11 Draining You may drain the capsule by application of air or nitrogen pressure (<1 bar |14.5 psi) to the inlet of the capsule. A dual air regulator system is recommended to prevent over-pressurization of the Sartobind capsule. -
Page 22: Operation Of The Sartobind Nano With Peristaltic Pumps Or Lc Systems
7.14 Operation of the Sartobind nano with peristaltic pumps or LC systems After the unit is filled completely with equilibration buffer; close the outlet of the Sartobind nano and remove the syringe. Start your LC system or peristaltic pump at a low flow rate. When fluid emerges, stop the pump, connect the tubing to the inlet of the Sartobind nano. - Page 23 Recommendations: – Maintain – Bed height (automatically kept constant when using capsules with same bed height) – Linear flow (automatically kept constant when scaling within capsules with same bed height) – Sample concentration – Increase (see scaling factors in the following table) –...
- Page 24 Capsule Factor to increase Factor to increase size height flow rate binding capacity [mL] [mm] (from nano) (from nano) – – 1200 5000 1667 1667 Example: After breakthrough experiments with the nano, you realize that a 1500-fold binding capacity is needed for a large scale run.
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Page 25: Testing Of Membrane Samples Enclosed With 5 L Capsules
8. Testing of membrane samples enclosed with 5 L capsules Membrane quality is always tested as described in chapter 10. Additionally the 5 L Jumbo is delivered with membrane samples of same membrane lot that was used in the capsule. This allows again for testing of membrane with user specific tests before use of capsule. -
Page 26: Method For Test Of Q Membrane Discs
Buffers: Equilibration buffer: 20 mM Tris |HCl, (pH 7.4, conductivity 1.80 mS/cm) Test solution: 0.2% BSA in equilibration buffer (2 mg/mL) Elution buffer: 1 M NaCl in equilibration buffer (Temperature of buffers: 20–25°C) Equipments: Glass or plastic dish pH meter Conductivity meter (temperature coefficient 2.00%/°C at 25°C reference temperature) Spectrophotometer set at 280 nm... - Page 27 Loading Apply 2 mL/cm test solution into the dish. Close the dish and shake for 12–18 h at 80 rpm. Remove excess test solution by vacuum pump as much as possible. Washing Apply 2 mL/cm of equilibration buffer and shake for 15 min. Remove equilibration buffer by vacuum pump.
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Page 28: Calculation Of Membrane Binding Capacity
8.1.3 Calculation of membrane binding capacity Binding capacity [mg/cm elute test elute + MA test + 2 + 10 elute 1.25 + 7.1 + 2.25 elute : extinction 280 nm of the elution solution elute : concentration of the test solution (2 mg/mL) test : volume of the elution solution (20 mL) elute... -
Page 29: Testing Sartobind S 30 Mm Discs
8.2 Testing Sartobind S 30 mm discs Using the appropriate conditions, lysozyme can be selectively bound to membrane adsorber 30 mm discs. By increasing the salt concentration of the buffer solution, the target substance can be recovered for subsequent quantification. Lysozyme in excess is loaded at a batch experiment on Sartobind S membrane disc. -
Page 30: Method For Test Of S Membrane Discs
Equipments: Plastic or glass dish pH meter Conductivity meter (temperature coefficient 2.00%/°C at 25°C reference temperature) Spectrophotometer set at 280 nm Orbital or reciprocal shaker set at approx. 80 rpm Operation temperature: 20–25°C 8.2.2 Method for test of S membrane discs Equilibration Apply 2 mL/cm equilibration buffer into the dish. - Page 31 Washing Apply 2 mL/cm of equilibration buffer and shake for 15 min. Remove equilibration buffer by vacuum pump. Repeat this step once. Elution Place each membrane disc in a separate Petri dish. Apply 10 mL elution buffer onto each membrane disc with a pipet. Shake for 1 h at 80 rpm (= elution solution).
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Page 32: Calculation Of Membrane Binding Capacity
8.2.3 Calculation of membrane binding capacity Binding capacity [mg/cm elute test elute + MA test + 2 + 10 elute 5.0 + 7.1 + 0.56 elute : extinction 280 nm of the elution solution elute : concentration of the test solution (2 mg/mL) test : volume of the elution solution (20 mL) elute... -
Page 33: Diffusion Test Of 75 Ml, 150 Ml, 1.2 L And
9. Diffusion test of 75 mL, 150 mL, 1.2 L and 5 L capsules The integrity can be controlled by a diffusion test. The testing procedure describes the diffusion test for pre and post use. The test is intended to discriminate between defective and intact capsules and to detect major bypasses, large holes and faulty assembly. - Page 34 Do not use pure water as this may lead to reversible swelling of the membrane strongly affecting the flow rate. Prior to integrity testing, the capsule needs to be prewashed, to remove any glycerol. The washing solution should be at room temperature. Keep the unit in an upright position for proper venting and open the vent screw on top of the unit until all air is replace by testing solvent.
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Page 35: Diffusion Measurement With Sartocheck 4
9.2.2 Diffusion measurement with Sartocheck ® 4 Plus Choose PROGRAMMING in the main menu Choose DIFFUSION TEST Test parameters Test pressure 200 mbar (2.9 psi) Stabilisiation time 2 min (75 mL capsule), 2 min (150 mL capsule), 3 min (1.2 L capsule), 5 min (Jumbo) Testing time 1 min... -
Page 36: Troubleshooting
10. Troubleshooting Problem Possible cause Action Air bubbles Incomplete air Small air bubbles seen in the top can be seen removal of the unit do not interfere with the purification as long as they do not touch the membrane bed. If too much air is enclosed, repeat removal as described in chapter 7.1 Venting. - Page 37 Problem Possible cause Action High back Sample has Prefilter with 0.2 µm filter before pressure not been processing through the unit. filtered Material has Proteins can form aggregates been filtered within hours or during operation. but was stored Thus we recommend to prefilter before purifica- inline by attaching a 0.2 µm filter tion...
- Page 38 Problem Possible cause Action Binding Process Use larger adsorber device, or: capacity is optimization connect two adsorbers (same size) not sufficient in series (i.e connect outlet of first adsorber to inlet of second) to achieve higher binding capacity. As a rule of thumb the pressure doubles when the flow rate is kept constant and the number of membrane layers is doubled.
- Page 39 Problem Possible cause Action Reuse is Laboratory The major application of capsules needed work is eased, is the single use and they are economic or constructed in plastic housing for practical this. Also they are validated and reasons certified only for one use. Technically they can be reused.
- Page 40 Problem Possible cause Action Binding Improper Prefilter with 0.2 µm filter before capacity filtration processing through the unit. decreases Some molecule Use capsule only once or use after several species binds DNA endonucleases such as uses ® Denarase tightly (e.g. DNA on Sartobind Q) and cannot be removed with...
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Page 41: Quality Assurance
Capsules and membranes are manufactured in a controlled environment. The product meets all Sartorius Stedim Biotech standards for traceability, production and specifications as given here or exceeded them as certified in the quality assurance certificate enclosed. -
Page 42: Ordering Information
12. Ordering information 12.1 Capsules Order number Description Quantity 96IEXQ42EUC11--A Sartobind Q nano 3 mL, Luer female connectors, manual, certificate 96IEXS42EUC11--A Sartobind S nano 3 mL, Luer female connectors, manual, certificate 96IEXQ42D9MFF--A Sartobind Q 75 mL, 3” sanitary clamp, manual, certificate 96IEXQ42D9MOO--A Sartobind Q 75 mL, hose barb connectors,... - Page 43 Order number Description Quantity 96IEXQ42E9BFF Sartobind Q 150 mL, 3” sanitary clamp, manual, certificate 96IEXS42E9BFF Sartobind S 150 mL, 3” sanitary clamp, manual, certificate 96IEXQ42E3FSS Sartobind Q 1.2 L, 11” sanitary clamp, manual, certificate 96IEXS42E3FSS Sartobind S 1.2 L, 11” sanitary clamp, manual, certificate 96IEXQ42E3ESS Sartobind Q jumbo 5 L,...
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Page 44: Accessories
12.2 Accessories Order number Description Quantity 5ZGl-0001 Stainless steel holder for 1 + 180 mL to 1.2 L Sartobind capsule 5ZGLG-0004 Stainless steel holder for 3 + 180 mL to 1.2 L Sartobind capsules 9ZGL--0102 Trolley for Jumbo 5 L, stainless steel 7ZAL-V0013 Reducing adapter... -
Page 45: Dimensions And Connections
13. Dimensions and connections Capsule size 3 mL 75 mL | 1.2 L 150 mL Dimensions 37 + 33 190 + 77 810 + 100 850 + 302 (height + diameter) Connectors Luer Sanitary Sanitary Sanitary female 11” 11” 3” 25 mm 50.5 mm 50.5 mm... - Page 46 Capsule size 3 mL 75 mL | 1.2 L 150 mL Recommended – 12.7 – – internal diameter of flexible tube Gaskets – 11”, 11”, 3”, inner inner inner diameter diameter diameter 16 mm 35.8 mm 35.8 mm...
- Page 48 Sartorius Stedim Biotech GmbH Copyright by Sartorius Stedim Biotech GmbH, Goettingen, Germany. August-Spindler-Strasse 11 37079 Goettingen, Germany All rights reserved. No part of this publication may be reprinted or Phone +49.551.308.0 translated in any form or by any means without the prior written Fax +49.551.308.3289...
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