Sartorius Stedim Biotech Sartobind Q Series User manual

Operating Instructions
Sartobind® Q and S
Membrane Adsorbers
Void Volume Optimized Capsules
With 4 | 8 mm Bed Height
85037-545-36
85037-545-36
nano 3 mL 75 mL | 150 mL 1.2 L Jumbo 5 L

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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. The membrane is dried from glycerol.
Die Verwendung dieser Produkte für Anwendungen,
für die sie nicht bestimmt oder nicht in dieser
Anleitung beschrieben sind, können zu einer
schlechteren Funktion, Zerstörung der Produkte
oder sogar zu Verletzungen von Mensch und Mate-
rial führen. Die Kapsulen sind nicht steril. Die ent-
haltene Membran wird aus Glycerin getrocknet.
L’utilisation des produits pour des applications non-
spécifiées ou décrites dans ce manuel peut causer
un disfonctionnement, une destruction du produit,
des dommages matériels ou même corporels. Les
capsules sont fournies non-stériles. La membrane
est séchée avec de la Glycérine.

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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
comportare un malfunzionamento, un danno al pro-
dotto stesso o a persone o cose. Le capsule sono
fornite non-sterilizzate. La membrana è asciugata
da glicerina.
본 설명서에 언급 또는 열거하지 않은 공정에서 본
제품을 사용하면 목적에 부합하지 않는 결과나, 인
체 상처 및 제품 또는 필터 재질에 손상을 초래할 수
있습니다. 캡슐필터는 비무균상태로 배송합니다.
필터의 여과막(멤브레인)은 글리세롤(glycerol) 로
부터 건조되었습니다.

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. Additional validation steps will be needed to
assure constant binding capacity and flow rate after each cycle.
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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.
Sartobind 75 and 150 mL capsules have been developed for
intermediate and pilot scale.
Sartobind 1.2 L and Jumbo 5 L capsules have been developed
for production purposes in the biopharmaceutical industry.
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1. Storage conditions ................................................................. 8
2. Introduction ............................................................................ 8
3. Technical data ....................................................................... 10
4. Materials ............................................................................... 12
5. Binding capacity ................................................................... 12
6. Installation ............................................................................ 13
7. Operation .............................................................................. 15
7.1 Venting ......................................................................... 15
7.2 Cleaning........................................................................ 16
7.3 Autoclaving .................................................................. 17
7.4 Recommended flow rates ............................................ 17
7.5 Buffer conditions ......................................................... 18
7.6 Selection of pH conditions .......................................... 19
7.7 Contaminant removal from therapeutic proteins
and other sources in flow-through mode .................. 19
7.8 Sample preparation ...................................................... 20
7.9 Washing ........................................................................ 20
7.10 Elution .......................................................................... 21
7.11 Draining ........................................................................ 21
7.12 Regeneration and storage ........................................... 21
7.13 Stability......................................................................... 21
7.14 Operation of the Sartobind nano with
peristaltic pumps or LC systems.................................. 22
7.15 Scaling up ..................................................................... 22
Table of Contents

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8. Testing of membrane samples enclosed
with 5 L capsules .................................................................. 25
8.1 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 .... 28
8.2 Testing Sartobind S 30 mm discs ............................... 29
8.2.1 Materials for test of S membrane discs ........... 29
8.2.2 Method for test of S membrane discs .............. 30
8.2.3 Calculation of membrane binding capacity .... 32
9. Diffusion test of 75 mL, 150 mL, 1.2 L and
5 liter capsules ...................................................................... 33
9.1 Installation for test ...................................................... 33
9.2 Operation of test .......................................................... 33
9.2.1 Pre-washing of capsule ..................................... 33
9.2.2 Diffusion measurement with Sartocheck® 4 .... 35
9.2.3 Results and evaluation ...................................... 35
10. Troubleshooting .................................................................... 36
11. Quality assurance ................................................................. 41
12. Ordering information ........................................................... 42
12.1 Capsules ........................................................................ 42
12.2 Accessories.................................................................... 44
13. Dimensions and connections ............................................... 45

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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.
They can be used for chromatographic separations in the down-
stream processing of viruses and proteins. The ion exchange
ligands are coupled to a membrane which is fitted into a plastic
housing for quick handling, making ion exchange purification
nearly as easy as filtration. The capsules are constructed with
optimized fluid channels. They carry a central core to minimize
void volume. Fluid models have been developed to reduce the
channel width to 3 mm. To set up and operate the Sartobind
Jumbo (5 liter bed volume) we recommend the Jumbo
trolley
(Fig. 4) available as accessory (order number 9ZGL--0102).
Capsules with strong basic and strong acidic ion exchange
Membrane Adsorber (MA) are available. These products are
intended for single use to avoid carryover as well as tedious and
1. Storage conditions

L
L
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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. The capsules
can also be used to capture large proteins, viruses such as adeno-
viruses and virus like particles (VLP).
Fig. 1: Sartobind nano 3 mL Fig. 2: Sartobind 75 and 150 mL,
arrows indicate the vent valves
Fig. 3: Jumbo 5 L (left) and 1.2 L Fig. 4: Jumbo 5 L on stainless, steel trolley
capsule, arrows indicate vent valves for Jumbo (accessory)
L
L
L
L

Membrane volume (MV) 3 mL 75 mL 150 mL 1.2 L 5 L
Nominal membrane area 110 cm22,700 cm25,500 cm244,000 cm2182,000 cm2
Bed height 8 mm 4 mm 8 mm 8 mm 8 mm
Design Cylindrical Cylindrical Cylindrical Cylindrical Cylindrical
Sartobind Q typical 10% dynamic binding capacity* 88 mg 2.16 g 4.4 g 35 g 145 g
Sartobind S typical 10% dynamic binding capacity* 77 mg 1.89 g 3.9 g 31 g 127 g
Maximum pressure bar (Mpa, psig) at 20°C 4 (0.4, 58) 4 (0.4, 58) 4 (0.4, 58) 4 (0.4, 58) 3 (0.3, 44)
Maximum pressure during venting
–
0.5 (0.05, 7) 0.5 (0.05, 7) 0.5 (0.05, 7) 0.5 (0.05, 7)
Nominal void volume 4 mL 180 mL 200 mL 1.6 L 7.0 L
Nominal void volume (MV) 1.3 2.4 1.3 1.3 1.4
Approximate weight 10 g 400 g 400 g 1.9 kg 16 kg | 20 kg wet |
23 kg filled
1 mL membrane = 36 cm2 membrane
Ion capacity per cm2 of membranes: 2–5 µeq
Short term pH stability Q | S: 1–14 | 3–14 refers to cleaning in place and regeneration
procedures during operation
Long term storage pH stability Q | S: 2–12 | 4–13 refers to overnight storage and longer.
Preferably store units in 20% ethanol | buffer within pH limits
* See also section 5. Binding capacity
3. Technical data
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Membrane volume (MV) 3 mL 75 mL 150 mL 1.2 L 5 L
Nominal membrane area 110 cm22,700 cm25,500 cm244,000 cm2182,000 cm2
Bed height 8 mm 4 mm 8 mm 8 mm 8 mm
Design Cylindrical Cylindrical Cylindrical Cylindrical Cylindrical
Sartobind Q typical 10% dynamic binding capacity* 88 mg 2.16 g 4.4 g 35 g 145 g
Sartobind S typical 10% dynamic binding capacity* 77 mg 1.89 g 3.9 g 31 g 127 g
Maximum pressure bar (Mpa, psig) at 20°C 4 (0.4, 58) 4 (0.4, 58) 4 (0.4, 58) 4 (0.4, 58) 3 (0.3, 44)
Maximum pressure during venting
–
0.5 (0.05, 7) 0.5 (0.05, 7) 0.5 (0.05, 7) 0.5 (0.05, 7)
Nominal void volume 4 mL 180 mL 200 mL 1.6 L 7.0 L
Nominal void volume (MV) 1.3 2.4 1.3 1.3 1.4
Approximate weight 10 g 400 g 400 g 1.9 kg 16 kg | 20 kg wet |
23 kg filled
1 mL membrane = 36 cm2 membrane
Ion capacity per cm2 of membranes: 2–5 µeq
Short term pH stability Q | S: 1–14 | 3–14 refers to cleaning in place and regeneration
procedures during operation
Long term storage pH stability Q | S: 2–12 | 4–13 refers to overnight storage and longer.
Preferably store units in 20% ethanol | buffer within pH limits
* See also section 5. Binding capacity
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Housing Polypropylene
Membrane matrix Stabilized reinforced cellulose nominal
pore size >3 µm
Ion exchange ligand Strong basic anion exchanger:
Sartobind Q quaternary ammonium
(R-CH2-N+(CH3)3)
Ion exchange ligand Strong acidic cation exchanger:
Sartobind S sulfonic acid
(R-CH2-SO3-)
5. Binding capacity
Data are based on dynamic binding capacity measurements
10% using 3 layers of 5 cm2 membrane discs (15 cm2 total area)
arranged in a holder and run at 10 mL/min.
Typical dynamic binding Reference protein
capacity 10% and buffer
Q 0.8 mg/cm2 (29 mg/mL) BSA (bovine serum albumin)
in 20 mM Tris/HCl, pH 7.5
S 0.7 mg/cm2 (25 mg/mL) Lysozyme in 10 mM
potassium phosphate, pH 7.0
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4. Materials

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.
Move the Jumbo trolley (accessory) in place. Then remove upper
foam protection and transparent bag. Lift the Jumbo directly
onto the trolley (inlet is up and the arrow imprinted on the ban-
derole is pointing down). We recommend to connect the Jumbo
with the trolley by the three screws delivered with the trolley.
The Jumbo 5 L carries protective caps on vent valves. To ensure
safe unpacking, the protective caps on inlet and outlet should
stay until you use the unit. Remove before venting. Store the
caps when you plan to autoclave (see chapter 7.3).
The capsules should be installed in an upright position in the
process flow. In this position the inlet is up. The flow is guided
to an external channel passing through the membrane layers to
an internal channel and to the outlet of the capsule.
Install the capsule in-line with a prefilter (0.2 µm) in front of the
device to prevent blockage or pressure built-up.
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6. Installation

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.
Fig. 5: Flow pattern inside the 150 mL capsule. 75 mL, 1.2 L and Jumbo 5 L are
constructed in the same way.
External channel
Membrane (4 | 8 mm)
Central core
Internal channel
The direction of
flow is imprinted
on the banderole of
the unit.
L
L
L
L
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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. After unpacking check vent valve
position. When turning left, the valve is open, when turning
right, the valve is closed. Before opening the vent valve, please
connect the valves with flexible tubing (inner diameter 6 mm)
to waste. During venting please do not exceed 0.05 MPa
(0.5 bar | 7.3 psi) pressure, as the vent valve O-ring could change
its position 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 nano fill a 10–20 mL Luer syringe with equilibration
buffer and connect to the capsule. Hold capsule upright (outlet
is up) and expel air as shown in Fig. 6. If you still detect any air
in the filled unit, close the outlet, hold the syringe up and move
the plunger slightly up and down that air bubbles can ascend
into the syringe. Another method is to connect a second empty
syringe to the top of the nano and expel air and buffer into that
syringe, disconnect the upper syringe to push out air and recon-
nect to the nano, turn it and purge the solvent back and and
forth. This method allows for observing bubbles in the unit.
7. Operation

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.
Fig. 6: Filling the Sartobind nano
with a Luer syringe
7.2 Cleaning
The modules have to be cleaned in place directly after unpacking
with 1 N NaOH, 30 min at 20°C.
1) 30 bed volumes (BV) 1 N NaOH solution at the flow rate of
1 BV/min at 20°C
2) 10 BV 1 N NaCl at 5 BV/min
3) 10 BV Equilibration buffer at 5 BV/min
Outlet
Inlet
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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. The protective caps enclosed in the
Jumbo delivery must be reinistalled on inlet and outlet connec-
tors of the Jumbo.
7.4 Recommended flow rates
Capsule Size Recommended Flow Rates (L/min)
3 mL 0.015
75 mL 1.5
150 mL 0.75
1.2 L 6
5 L 25
As a rule of thumb, flow rates of 5 membrane volumes per
minute are recommended for 8 mm bed height and 10–30 mem-
brane volumes for 4 mm bed height. Lower flow rates than the
recommended ones can also be used but will typically not
improve binding capacity or overall performance.

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.
The buffer should have a pKa within 0.5 pH units of the pH
used. It should be filtered with 0.2 µm filters before use and the
quality of water and chemicals should be of high purity.
Do not apply pure water as it may lead to a
reversible swelling and decrease of the flow rate of
the membrane. The buffering ion should carry the
same charge as the ion exchange ligand.
The minimum buffer volume for equilibration is the
same as the flushing volume.
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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. In buffers below the
isoelectric point (at least 1 pH unit), a protein, for example car-
ries a positive net charge and will bind to a cation exchanger
(Sartobind S). In buffers above its isoelectric point (at least 1 pH
unit), it will bind to an anion exchanger (Sartobind Q).
7.7 Contaminant removal from therapeutic proteins
and other sources in flow-through mode
For contaminant removal from products such as mono clonal
antibodies, pH conditions in the range of pH 6 to 8 are used
in order to bind highly negatively charged DNA, endotoxins,
contaminating proteins, some host cell proteins and viruses.
The product of interest, the monoclonal antibody with
isoelectric points (pI) of 8 – 9.5 for example, will not bind and
pass through Sartobind Q.

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.
7.8 Sample preparation
The sample should be adjusted to the starting buffer
conditions and be prefiltered through a 0.2 µm membrane filter
e.g. Sartopore 2 capsule (order number 544307H1). For small
volumes in the mL range use a 0.2 µm Minisart® filter with Luer
outlet (order number 16534-K).
Unfiltered feed will block the Membranes Adsorber
and lead to capacity loss and increased back
pressure. We recommend inline filtering during
operation. With increase of pressure replace filter
and restart.
7.9 Washing
When using capsules in bind & elute mode, wash with equilibra-
tion buffer after sample loading.
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