INCREASE® Application Support

Crossflow Filtration – SartoJet with Sartocon Slice

Standard Operating Procedures

Prepared for Prepared by

Mr Seow Tan Ze Yu
Application Engineer
Prime Biologics Pte Ltd
Purification Technologies
41 Science Park Road,
#01-09, The Gemini (Lobby A)
www.sartorius-stedim.com
Singapore 117610
Contents

1 Purpose .................................................................................................................................................... 3

2 Equipment and Materials.................................................................................................................... 3

2.1 Equipment .................................................................................................................................................................................... 3

2.2 Material ......................................................................................................................................................................................... 4
2.3 Set-Up............................................................................................................................................................................................ 5

3 Procedure ................................................................................................................................................ 5

3.1 Setting up ..................................................................................................................................................................................... 5

3.2 Flushing the filter cassettes .................................................................................................................................................... 6
3.3 Determining the water flux rate for filter cassettes (CWF) .......................................................................................... 6
3.4 Measure the hold-up volume in the SartoJet system .................................................................................................... 6
3.5 Crossflow Ultrafiltration Process .......................................................................................................................................... 7
3.6 Cleaning the filter cassettes ................................................................................................................................................... 7
3.7 Storing the filter cassettes ...................................................................................................................................................... 8
3.8 IPA Wipe Down of System ...................................................................................................................................................... 8

4 Integrity Test .......................................................................................................................................... 8

4.1 Manual Integrity Test................................................................................................................................................................ 9

4.2 Automatic Integrity Test ........................................................................................................................................................10
4.3 Integrity Test Values ................................................................................................................................................................11

5 Additional Support ............................................................................................................................. 11

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 2|11

1 Purpose
The purpose of this document is to outline the procedures of carrying out ultrafiltration process
steps using the Sartocon Slice holder with the SartoJet pump.

2 Equipment and Materials

2.1 Equipment
1) SartoJet Pump

Figure 1 – Picture of SartoJet Pump

2) Sartocon Slice 0.1 m2 Cassettes

Figure 2 – Picture of Sartocon Slice 0.1m2 Cassettes

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 3|11

 3) Sartocon Slice Stainless Steel Holder (with all pressure gauges and valves)

Figure 3 – Picture of Sartocon Slice Holder

4) Silicon Tubing
5) Torque wrench
6) Weighing Balance
7) pH indicator
8) Measuring Cylinder
9) Hose barb adapters
10) ¾” Tri-clamps and EPDM gaskets

2.2 Material
1) Sample
2) WFI
3) 5 L of 1M Sodium hydroxide
4) 0.1M Sodium hydroxide
5) 20% Ethanol
6) IPA

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 4|11

 2.3 Set-Up
Set up the equipment according to the schematic diagram below.

Retentate side
PI-1
TFF

V-1

PI-2 PI-3
Sartorius
SartoJet®

V-2
Feed Side Permeate Side
0.000g
Sartorius Balance WB-01
P-01

Figure 4 – Schematic Diagram of equipment set-up

3 Procedure

3.1 Setting up
1) Remove filter cassette from its packaging and handle the filter by the outer edges only.
2) Check and record cassette from its packaging.
3) Check and record cassette labelling.
4) Place the filter cassette with the semi-circular cut-outs facing downward on the two
guide rods between the two holding plates.
5) Tighten alternately the nuts until the filter cassette is secured in place by the holding
plates.
6) Use a torque wrench to tighten the nuts in opposite diagonal pairs to the torque
specified below
i. Sartocon Slice holding device: 20 Nm for Hydrosart cassettes
25 Nm for PES cassettes
ii. Do not exceed, even briefly, the permissible torque, check the hydraulic pressure
or torque before use.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 5|11

3.2 Flushing the filter cassettes
7) The cassette must be flushed with 10 L WFI per 0.1 m2 filter area at operating condition:
i. Pin = 1.0 bar
ii. Pret = 0.0 bar
iii. Pper = 0.0 bar (closed for the first 10 minutes)

8) Open the all the valves and start the pump.

9) Allow both the retentate line and permeate line to be drained by pumping air through
the system.

3.3 Determining the water flux rate for filter cassettes (CWF)
The purpose of comparing the initial flux rate with the flux rate determined after every cleaning
procedure will enable the evaluation of how well the cassette has been cleaned.
CWF is determined by measuring the permeate flow rate when flushed with water at a particular
operating condition.

Table 1 – Water flux rate evaluation

Measurement Pin Pret Pper TMP T CWF

before filtration (bar) (bar) (bar) (bar)
o
( C) (LPM)
1 1 0 0 0.5 25 IF1
2 2 0.5 0 1.25 25 IF2

Measurement after Pin Pret Pper TMP T CWF

CIP (bar) (bar) (bar) (bar)
o
( C) (LPM)
1 1 0 0 0.5 25 FF1
2 2 0.5 0 1.25 25 FF2

Recovery % : 1 IF1 / FF1

2 IF2 / FF2

3.4 Measure the hold-up volume in the SartoJet system

10) Load the sample into the feed tank and measure the initial weight of the sample.
11) Close the permeate valve and open the retentate valve and allow for recirculation of the
sample in the SartoJet system.
12) Stop the pump and measure the new weight.
13) Hold-up volume = Initial weight – Final weight.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 6|11

3.5 Crossflow Ultrafiltration Process
14) Close the permeate valve, open the retentate valve and start the pump.
15) Slowly open the permeate valve till full.
16) Adjust the retentate valve and pump efficiency till the desired operating conditions.
17) Continue concentrating till the desired concentration factors while recording the time,
concentration factor and flux at regular permeate volumes.

Table 2 – Data recording table during concentration

Permeate
Time Inlet Outlet TMP Conc.
Volume
min bar bar bar bar Litre Factor

18) Use either buffer solution or air to recover the hold-up volume in the system.

3.6 Cleaning the filter cassettes

19) Direct both retentate and permeate lines to a waste container. Flush the filter cassettes
with 10 L WFI for at room temperature.
Conditions:
pin = 1.0 bar
pret = 0.0 bar
pper = 0.0 bar
20) Flush 1 L of 1M NaOH with the same conditions as above. Stop the pump upon
completion.
21) Direct the retentate line back to the NaOH feed tank. Recirculate the remaining NaOH
for 60 minutes; then open the permeate valve and flush the filter cassettes with the
same batch of NaOH until all the NaOH has been flushed out from the permeate line.
Stop the pump upon completion.
Conditions:
pin = 1 bar
pret = 0 bar
pper = closed (opened after 60 min)

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 7|11

 22) Direct the retentate line back to the waste container. Flush with purified water until the
pH is neutral (check with pH strips).
Conditions:
pin = 1 bar
pret = 0 bar
pper = 0 bar
23) Measure the water flux recovery and complete the table at section 3.2.1.
If Recovery % is less than 90%, repeat steps 22 and 23. If required, elevate operating
conditions to achieve better cleaning such as higher inlet pressure, running step 22 for a
longer time, using a different cleaning agent, or elevate the temperature of the cleaning
agent.

3.7 Storing the filter cassettes

24) Remove the filter cassettes from the holding device and store the filter cassettes in a
either 20% ethanol or 0.1M NaOH in a closed container or bag, ensuring that the filter
cassettes are completely submerged in the solution.
25) For storing of cassettes in the holder:
Direct the retentate line back to a feed tank with 0.1M of NaOH. Recirculate the 0.1 M
of NaOH for 5 minutes at the same flow ratio that you have already used for flushing
the filter cassettes. Stop the pump and close the feed inlet, retentate outlet and
permeate outlet. Do not allow cassesttes to dry out or freeze.
Execute step 22 before the next run.

3.8 IPA Wipe Down of System

26) Perform a simple IPA wipe down for all exposed parts.

4 Integrity Test
Before conducting Integrity test, ensure that the filter cassettes are properly wetted by following
the steps outlined in section 3.2. The air diffusion rates given in Integrity Test Values are
reference values only.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 8|11

4.1 Manual Integrity Test
27) Attach a hose to the feed inlet (V1) and connect the other end of the hose to a
compressed air source.
28) Close the retentate outlet (V2).
29) Adjust the inlet pressure P1 to the value specified in the Integrity Test Values (see below
in section 4.3) and keep this pressure constant.
30) At a constant inlet pressure P1, wait for four minutes until the system has stabilized
31) Use a flow meter or an inverted burette filled with water to measure the air diffusion at
the permeate side.

Figure 5 – System set up for manual integrity testing

* It does not matter if the air source is connected to the feed or retentate side. If the air source is
connected to the retentate side, the feed side will be closed and vice versa.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 9|11

4.2 Automatic Integrity Test
32) Connect an integrity tester (e.g. Sartocheck® 4plus) to the power supply and pressure
source.
33) Connect the integrity tester “Outlet line” to the retentate line.
34) Close the feed line and open the permeate line.
35) Enter the required test parameter according to the integrity tester instruction manual
and cassette data
36) Use the following parameters:
Stabilization time: 4 minutes
Test time: 2 minutes
37) Enter the maximum allowable diffusion value (see Integrity Test Values below in section
4.3)

Figure 6 – System set up for automatic integrity testing

* It does not matter if the integrity tester is connected to the feed or retentate side. If the
integrity tester is connected to the feed side, the retentate side will be closed and vice versa.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 10|11

4.3 Integrity Test Values

If the measured air diffusion rates exceed those values, the membrane might be damaged.
High diffusion values can be caused by incomplete wetting of membranes or insufficient
torque pressure.

5 Additional Support
For more information and application support, do not hesitate to contact your local Sartorius
Stedim Biotech application specialists.

INCREASE ® SOP–TFF-13-004 Updated: Dec-2014 ZY Tan 11|11