Sartorius Sartobind IEX MA 15,Sartobind IEX MA 75,Sartobind IEX MA 100,Sartobind IEX MA Series Operating Instructions Manual
Operating Instructions
Sartobind
®
IEX MA 15 | 75 |100
A Separation Technology
Based on Macroporous Membranes
85032-539-13
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.
The units are supplied as non-sterile.
The membrane is dried from glycerol and should be prewashed with
equilibration buffer before use.
If you plan to scale up later, preferentially use Sartobind capsules
with 4 or 8 mm bed height: e.g. Sartobind Q nano with 1 or 3 ml as
downscale.
2
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.
Sartobind MA 100 units with 5 membrane layers can be used for
screening of buffer conditions and quantitative work at exceptional
high flow rate.
3
4
1. Storage conditions 5
2. Introduction 5
3. Technical data 7
4. Materials and pH
stability 8
5. Binding capacity 9
6. Installation 10
7. Operation 11
7.1 Venting 11
7.2 Recommended buffer
volumes and flow rates 11
7.3 Buffer conditons 13
7.4
Selection of
pH conditions
14
7.5 Contaminant removal
from proteins in flow
through operation 14
Table of Contents
7.6 Sample preparation
and equilibration 15
7.7 Washing 16
7.8 Elution 16
7.9 Regeneration,
cleaning and storage 17
7.10 Stability 18
7.11 Operation with
peristaltic pump or
LC system 19
8. Troubleshooting 20
9. Quality assurance 24
10. Ordering information 25
10.1 MA units 25
10.2 Accessories 26
11. Dimensions and
connections 27
5
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. The limiting factor in
low pressure column chromatography is the time required for the
molecules to diffuse into and out of the pores. The various steps of
equilibration, loading, washing, elution and regeneration can take
hours.
Sartobind membranes are made from regenerated and stabilized cellulose. The stabilization and cross-linking generate high chemical
stability. Conventional ion exchange ligands are covalently attached
to the macroporous membrane support. The membrane layers form
chromatographic bed between 3 and 15 layers which is incorporated
into housings.
1. Storage conditions
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 – 30 fold compared to traditional column chromatography.
The Luer-Lok connectors on top and bottom allow you to run the
units with a syringe and a prefilter or to connect several units to
each other to get a larger bed volume or to run units with different
chemistries.
6
7
MA 15 MA 75 MA 100
Membrane Area [cm2] 15 75 100
Number of layers 3 15 5
Bed height [mm] 0.8 4.0 1.4
Membrane volume [ml] 0.41 2.1 2.8
Membrane diameter [mm] 25 25 50
Typical dynamic binding 12 60 80
capacity 10% for Q [mg/unit]*
Typical dynamic binding 10.5 52.5 70
capacity 10% for S [mg/unit]*
Typical dynamic binding capacity – 60 –
10% for D [mg/unit]**
Ligand density [μeq/cm2] 2 – 5 (Q, S), 2 – 4 (D)**
Maximum pressure at 20°C 6 bar | 0.6 MPa | 87 psi
* See also section 5. Binding capacity
** D type is available as MA 75 only.
3. Technical data
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-CH2–N+(CH3)
3
Strong acidic cation exchanger Sulfonic acid (S)
R-CH2–SO
3
–
Weak basic anion exchanger Diethylamine (D)
R-CH
2
–N(C2H5)
2
8
4. Materials
9
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/cm2) and buffer
Q 0.8 BSA (bovine serum
albumin) in 20 mM
Tris/HCl, pH 7.5
S 0.7 Lysozyme in 10 mM
potassium phosphate,
pH 7.0
D 0.8 BSA (bovine serum
albumin) in 20 mM
Bis Tris, pH 6.0
5. Binding capacityy
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