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Summary of Contents for Sartorius Sartobind IEX MA 15
- Page 1 Operating Instructions ® Sartobind IEX MA 15 | 75 |100 A Separation Technology Based on Macroporous Membranes 85032-539-13...
- Page 2 Read operational instructions carefully before using Sartobind capsules. ! Important Use of the product in applications not specified or not described in this manual, may result in improper function, personal injury, or damage of the product or material. Membrane Adsorber (MA) units should be visually inspected before use.
- Page 3 Intended use The products are intended for ion exchange (IEX) chromatography work in a laboratory for research purposes only. Sartobind MA 15 units with 3 membrane layers can be used for screening of buffer conditions and quantitative purifications at exceptional high flow rate. Sartobind MA 75 units contain 15 membrane layers and have been developed for bind and elute and flowthrough applications at high flow rate.
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Page 4: Table Of Contents
Table of Contents Storage conditions 7.6 Sample preparation and equilibration Introduction 7.7 Washing 7.8 Elution Technical data 7.9 Regeneration, Materials and pH cleaning and storage stability 7.10 Stability Binding capacity 7.11 Operation with peristaltic pump or Installation LC system Operation Troubleshooting 7.1 Venting 7.2 Recommended buffer... -
Page 5: Storage Conditions
1. Storage conditions Sartobind MA units should be stored clean, dry and away from direct sunlight in the box at room temperature. 2. Introduction Traditional chromatography uses porous particles packed into columns. As liquid flows through the column and around the beads, bio-molecules in the liquid diffuse into the pores of the beads to binding sites on the inner surface of the pores. - Page 6 Pressure forces the liquid through the pores of the membrane, bringing target substances to direct contact with the binding sites. This direct convection to the binding sites minimizes diffusion limitation of mass transfer and increases the speed of operation to 10 –...
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Page 7: Technical Data
3. Technical data MA 15 MA 75 MA 100 Membrane Area [cm Number of layers Bed height [mm] Membrane volume [ml] 0.41 Membrane diameter [mm] Typical dynamic binding capacity 10% for Q [mg/unit]* Typical dynamic binding 10.5 52.5 capacity 10% for S [mg/unit]* Typical dynamic binding capacity –... - Page 8 4. Materials Housing and basic membrane Housing MA 15, 75, 100 Polysulfone Membrane matrix Stabilized reinforced cellulose, nominal pore size >3 μm Membrane ligands Quaternary ammonium (Q) Strong basic anion exchanger R-CH –N Strong acidic cation exchanger Sulfonic acid (S) –...
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Page 9: Binding Capacity
5. Binding capacityy Data are based on dynamic binding capacity measurements 10% using MA15 run at 10 ml/min. Membrane Typical dynamic binding Reference protein capacity 10% (mg/cm and buffer BSA (bovine serum albumin) in 20 mM Tris/HCl, pH 7.5 Lysozyme in 10 mM potassium phosphate, pH 7.0 BSA (bovine serum... -
Page 10: Installation
6. Installation The contents of the packages are described in chapter 10.1. The Sartobind Membrane Adsorber (MA) units are ready-to-use devices and can be used out of the box. They can be operated with a syringe (chapter 7.1 to 7.8), peristaltic pump or liquid chromatogra- phy (LC) system (follow chapter 7.11 first). -
Page 11: Operation
7. Operation 7.1 Venting It is important to remove air from the unit completely. Fill a 10 – 20 ml Luer syringe with equilibration buffer and connect to Luer female inlet, then hold unit upright (outlet is up) and expel air until the first fluid is seen at the outlet. - Page 12 Tab. 1: Buffer consumption of Sartobind MA units [ml] MA 15 MA 75 MA 100 Equilibration Wash 10 – 20 10 – 20 10 – 20 Elution 5 – 20 5 – 20 5 – 20 Regeneration 5 – 10 10 –...
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Page 13: Buffer Conditons
7.3 Buffer conditions In the majority of applications an equilibration buffer concentration 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. The buffer should have a pKa within 0.5 pH units of the pH used. -
Page 14: Selection Of Ph Conditions
7.4 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 elution buffer. The pH value at which a biomolecule has no net charge is the isoelectric point (pI). -
Page 15: Sample Preparation And Equilibration
To remove contaminating proteins and aggregates with Sartobind S in flow through mode, process impurities have to be charged posi- tively to bind while the target protein stays negative. At the pH of the buffer above the pI, the protein product flows through without binding. -
Page 16: Washing
The Minisart shall be coupled directly to inlet of the MA unit during use as shown in Fig. 2 to avoid clogging of the adsorber. This is recommended also for usage of MA units with LC systems. ! Important Unfiltered feed will block the Membrane Adsorber and lead to capacity loss and increased back pressure. -
Page 17: Regeneration, Cleaning And Storage
7.9 Regeneration, cleaning and storage After elution, wash the unit with equilibration buffer. If necessary, use 1 N NaOH, 1 N HCl or 70% ethanol for 1 hour and store in 20% ethanol in equilibration buffer. Do not store units in water. Do not freeze the units. -
Page 18: Stability
7.10 Stability The MA 15, 75 and 100 units are stable against all commonly used buffers in chromatography, e.g. 8 M urea, 8 M guanidine hydrochlo- ride, ethanol and acetone. They can be sanitized after prewashing with 1 N NaOH, 1 hour at room temperature (20°C). Do not use oxidizing agents such as hypochlorite or H pH Stability Short term*... -
Page 19: Operation With Peristaltic Pump Or Lc System
7.11 Operation with peristaltic pump or LC system After the unit is filled completely with equilibration buffer, close the outlet of the MA 15, 75 or 100 and remove the syringe. Start your LC system or peristaltic pump at a low flow rate. When fluid emerges, stop the pump and connect the tubing to the inlet of the unit. -
Page 20: Troubleshooting
8. Troubleshooting Problem Possible cause Action High back Material has Prefilter with 0.2 μm filter pressure not been before processing through the during sample filtered unit. loading Material has Proteins can form aggregates been filtered within hours or during but was stored operation. Thus we recommend before to prefilter inline by attaching a purification... - Page 21 Problem Possible cause Action Target mole- Conditions for Decrease conductivity, control other cule is not binding are process parameters as type of buffer bound insufficient and pH Proteins or Perform a regeneration cycle contaminants are still bound from last cycle Binding capa- Improper Prefilter with 0.2 μm filter before city decreases filtration...
- Page 22 Problem Possible cause Action Proteins or Run a 1 M NaCl buffer step to contaminants elute tightly bound proteins are still bound quantitatively. Then regenerate from last cycle adsorber by loading with 1 N NaOH and keep it for 1 hour at room temperature (20°C).
- Page 23 Problem Possible cause Action Early Binding Use larger adsorber device, or break- capacity is connect two adsorbers (same size) through of not sufficient in series (i.e. connect outlet of first protein 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...
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Page 24: Quality Assurance
Sartobind MA units are manufactured in a controlled environment and have been tested for protein binding capacity and flow rate. The product meets all Sartorius Stedim Biotech standards for traceability, production and specifications as given here or exceeded them. -
Page 25: Ordering Information
10. Ordering information 10.1 MA units Order number Description Quantity Sartobind MA units (reusable) 93IEXQ42GB-12--A Sartobind Q 15, Luer female and male connectors 93IEXS42GB-12--A Sartobind S 15 Luer female and male connectors 93IEXQ42DB-12--V Sartobind Q 75, Luer female and male connectors 93IEXS42DB-12--V Sartobind S 75, Luer female and male connectors... -
Page 26: Accessories
10.2 Accessories Order number Description Quantity 1ZA- - -0004 Luer male adapter to UNF 10-32 female, PEEK 1ZA- - -0005 Luer female adapter to UNF 10-32 female, PEEK 17002---140 2 + M6 female to female Luer and 2 + M6 female to male Luer Lok Adapter, black Tefzel... -
Page 27: Dimensions And Connections
11. Dimensions and Connections MA 15 MA 75 MA 100 Membrane Area [cm Membrane volume [ml] 0.41 Dimensions 25 + 36 28 + 36 31 + 66 (height + diameter) [mm] Frontal surface area [cm Connector inlet Luer female Connector outlet Luer male Approximate weight [g]... - Page 28 Luer-Lok is a trademark of Becton, Dickinson and Company. Printed in Germany on paper that Published has been bleached without any use 28. February, 2011 of chlorine Sartorius Stedim Biotech GmbH, W4A000 Sartobind IEX MA 15|75|100 Goettingen, Germany Publication No.: SL-6171-e11011 Ver. 02 | 2011...
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