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Continuous Capture Chromatography
A simple, robust approach to move from batch to
continuous capture
LEWA Bioprocess Technologies Group – Devens, MA USA
Contents
– Our approach to multi-column capture chromatography
LEWA-Nikkiso America Inc.
2
– Customer case studies
– Conclusions
– A simple set up
– Who we are and what we do
Why use continuous chromatography for capture?
Under utilization of resin capacity in Protein A step
• Protein A resin costs: $10,000 – $15,000 / L
• Size of Protein A columns used in manufacturing: 100 – 250 L
• One packed column worth $1 – $2.5 million, but utilization is only 40-60%!
© LEWA - Bioprocess Group and 3
Unused resin capacity,
typically 40-60% Feed
(clarified harvest)
Batch single Protein A column Flow-through
4© LEWA - Bioprocess Group
Why continuous chromatography for capture?
The driver for the development of continuous capture processes using counter-current
chromatography was the realization that 40 – 60% of the costly Protein A resin was not being utilized.
Even though people started thinking about the under utilization of Protein A resin back in the mid-
2000s, developing improvements didn’t become a priority until processes went to large scale
production where the cost of one Protein A column could be in excess of several millions of dollars.
The basic principle of continuous capture (2C-PCC*)
5
feed
unused resin
capacity
feed
fully loaded column
elution
*2C-PCC: 2-Column Periodic Countercurrent Chromatography© LEWA - Bioprocess Group and
6© LEWA - Bioprocess Group
The basic principle of continuous capture (2C-PCC*)
Different terminology such as, “periodic countercurrent chromatography,” “multi-column continuous
capture chromatography” generally refer to the same technology.
In continuous capture using two columns, the first column is fully loaded and breaks through to the
second column, then the second column is fully loaded so that all resin capacity is utilized. After the
initial loads, while one column is being loaded, the other column is washed, eluted, regenerated, and
re-equilibrated. This cycle is repeated continuously until the entire batch has been processed.
ChromaCon / LEWA Partnership
LEWA is global GMP scale up partner for
ChromaCon two column technology.
Control logic for operating two columns
continuously on a single system.
7
ChromaCon
•Process know-how
•Intellectual property
•Entry-stage equipment
LEWA
•Scale-up Equipment
•Process solutions
•Continuous manufacturing
•Worldwide Footprint
CaptureSMB
MCSGP
• Technical, marketing and sales
collaboration.
• Patented technology
ChromaCon / LEWA Partnership
LEWA is global GMP scale up partner for
ChromaCon two column technology.
Control logic for operating two columns
continuously on a single system.
8
CaptureSMB
MCSGP
• Technical, marketing and sales
collaboration.
• Patented technology
The continuous capture chromatography that we employ in our GMP scale system is based on and
licensed from ChromaCon, a Swiss life science tool company that specializes in developing and
commercializing chromatographic process technology for protein and monoclonal antibodies.
Continuous capture with LEWA EcoPrime® Twin LPLC
© LEWA Bioprocess Group and 9
Huge productivity10 cm ID Protein A columns –
more than 1 kg mAb purified/day
(500 L feed)
SimplicityTwo column design minimizes
complexity and secures
maximum uptime
Easy scale upProcess design on the
Contichrom CUBE, fast scale-up
to EcoPrime Twin
Continuous capture with LEWA EcoPrime® Twin LPLC
© LEWA Bioprocess Group and 10
There are 3 main advantages for using EcoPrime Twin LPLC for continuous capture.
1. Significant increase in productivity as compared with traditional batch operation. For example,
over 1 kg of mAb can easily be purified in a day on a Twin with two 10-cm ID Protein A columns.
2. The simple 2-column design of the EcoPrime Twin dramatically reduces the complexity when
compared to systems with 3 or more columns. This simplicity minimizes downtime and facilitates
transfer to manufacturing and validation.
3. The EcoPrime Twin was designed to be the scale-up system for the Contichrom CUBE, a bench
scale system. Methods from the CUBE can be easily translated for configuration and operation on
the Twin.
Huge productivity
System/ModelColumn ID Range
(cm)
Feed per day
(L/day)
Product recovered*
(kg/day)
EP Twin 1000.004 – 0.60 L/min
2.5 10 23 360 0.11 1.7
EP Twin 2500.02 – 2.4 L/min
5 20 92 1,460 0.44 7.0
EP Twin 5000.06 – 9.0 L/min
8 45 206 7,420 1.0 35.0
© LEWA Bioprocess Group and 11
10 cm ID columns
More than 300 L feed processed/day
More than 1 kg mAb purified/day
*Process dependent
Huge productivity
System/ModelColumn ID Range
(cm)
Feed per day
(L/day)
Product recovered*
(kg/day)
EP Twin 1000.004 – 0.60 L/min
2.5 10 23 360 0.11 1.7
EP Twin 2500.02 – 2.4 L/min
5 20 92 1,460 0.44 7.0
EP Twin 5000.06 – 9.0 L/min
8 45 206 7,420 1.0 35.0
© LEWA Bioprocess Group and 12
10 cm ID columns
More than 300 L feed processed/day
More than 1 kg mAb purified/day
*Process dependent
Modeling parameters:
Linear velocity feed (cm/h) – 250
Linear velocity process(cm/h) – 300
Step yield (%) – 95
Titer (g/L) – 5
Column length – 10
DBC (g/L resin) – 60
Productivity calculated – 1.11 kg/L resin/ day
Buffer consumption calculated – 0.41 L/g product
The numbers in the table are estimates of the productivity that can be achieved with the EcoPrime
Twin. The EcoPrime Twin is available in 3 sizes based on the flow rate capability. Even with the
smallest Twin and 10-cm ID columns, over 300L of feed can be processed in a day generated 1.7 kg
of product.
Simplicity
The 2-column approach reduces risk
• Accelerates validation
• Simplifies maintenance
• Lessens costly downtime
• Minimizes operating expenses
• Streamlines implementation, easy-to-
understand and use
© LEWA - Bioprocess Group and 13
8 column multicolumn process
~240 Valves
~40 Valves
Same productivity as systems with 3 or
more columns, in a simple 2-column design.
CUBE from ChromaCon®
Simplicity
The 2-column approach reduces risk
• Accelerates validation
• Simplifies maintenance
• Lessens costly downtime
• Minimizes operating expenses
• Streamlines implementation, easy-to-
understand and use
© LEWA - Bioprocess Group and 14
8 column multicolumn process
~240 Valves
~40 Valves
CUBE from ChromaCon®
A key element of the simplicity inherent in the 2-column design of the EcoPrime Twin and the CUBE is
the significant reduction in valves required. Fewer mechanical components not only means less
downtime for repair and maintenance, but also accelerates validation and streamlines implementation
in a manufacturing setting because the equipment and it’s operation is so much easier to understand.
The real beauty of the simple, 2-column design is that we can achieve the same productivity as
systems with 3 or more columns.
Easy scale-up, scale-down
© LEWA - Bioprocess Group and 15
Contichrom CUBE EcoPrime Twin 100 EcoPrime Twin 500
CUBE is a simple tool for scaling-up and scaling down.• Develop new processes
• Troubleshoot existing processes
EcoPrime Twin mirrors CUBE operation with continuous capture, batch, and
sequential batch functionality.
Easy scale-up, scale-down
© LEWA - Bioprocess Group and 16
Contichrom CUBE EcoPrime Twin 100 EcoPrime Twin 500
CUBE is a simple tool for scaling-up and scaling down.• Develop new processes
• Troubleshoot existing processes
Whether you are developing new processes or troubleshooting an existing one, scaling up to the Twin
or down to CUBE is relatively easy because the design of the Twin mirrors the operation of the CUBE.
There is nothing worse for a schedule than having to redo your process development when scaling
up. Or, when things go wrong, not have a scaled-down system to do quick troubleshooting on.
UV signals of four cycles (column 1 and column 2)
Seamless scale-up of a continuous capture process
17
Contichrom
CUBE
© LEWA - Bioprocess Group and
EcoPrime
Twin
UV signals of four cycles (column 1 and column 2)
Seamless scale-up of a continuous capture process
18
Contichrom
CUBE
© LEWA - Bioprocess Group and
EcoPrime
Twin
Shown on this slide is an example of what you can expect when scaling up to the Twin from the
CUBE. The transfer of the method operation is seamless and reproducible.
Contents
– Our approach to multi-column capture chromatography
LEWA-Nikkiso America Inc.
19
– Customer case studies
– Conclusions
– A simple set up
– Who we are and what we do
20© LEWA - Bioprocess Group
Batch Capture Total CV 7.85 L
Continuous Capture Total CV 1.6 L
Simple transfer from batch to continuous capture
Customer A - process development run comparison with 1g/L titer mAb
Essentially same process steps = simple process transfer
Higher load volume 40 vs 49 CVs = higher resin utilization & reduce buffer
consumption
Shorter residence times = faster loading & shorter processing time
Linear
[cm/h]
Column Equilibration 3 250
Load 40 150
Wash 1 2 150
Wash 2 2 250
Wash 3 3 250
Elution 3 250
post wash 1 3 250
post wash 2* 2 250
Regen* 3 250
CV
Process time: 9 h Cycle time: 4 h at linear velocity of 250 cm/h
CV
Load Start-Up 15
Load Connected 27
Load Parallel 22
Wash 1 connected 1
Wash 1 Parallel 1
Wash 2 2
Wash 3 3
Elution 3
Post-Wash1 2
Post Wash 2 1
Regeneration 3
Re-Equilibration 5
21© LEWA - Bioprocess Group
Batch Capture Total CV 7.85 L
Continuous Capture Total CV 1.6 L
Simple transfer from batch to continuous capture
Customer A - process development run comparison with 1g/L titer mAb
Process time: 9 h Cycle time: 4 h at linear velocity of 250 cm/h
In the first case study, the customer transferred their traditional mAb purification platform batch
capture step to a continuous capture step on the LEWA EcoPrime Twin for a 1 g/L titer mAb.
Essentially the same steps are employed when transferring from a batch process to a continuous
capture process so the method or recipe transfer is easy and straightforward. You will notice that with
the continuous capture method, higher load volumes are possible (49 CV and compared to 40 CV)
since the breakthrough from one column on the second column. The result is higher resin utilization
and reduced buffer consumption. In addition, because the breakthrough can be captured on the
second column, less residence time is needed when loading the column, so faster loading results in
shorter processing time.
22© LEWA - Bioprocess Group
EcoPrime Twin continuous capture reproducibility
Customer A –1g/L titer mAb: 50 g/L load
• At steady state, variability of peak area is <1 %.
• Steady state can be reached with first elution.
Column 1 Column 2
1 0.250 0.294
2 0.263 0.297
3 0.265 0.297
4 0.265 0.297
5 0.265 0.297
Average 0.264 0.297
error %* 0.639 0.025
SD 0.0064 0.0014
AreaPeak
5 peaks overlaid
Column 1
Column 2
23© LEWA - Bioprocess Group
EcoPrime Twin continuous capture reproducibility
Customer A –1g/L titer mAb: 50 g/L load 5 peaks overlaid
Column 1
Column 2
The data on this slide demonstrates the reproducibility of five cycles of a continuous capture run with 1
g/L mAb. A load of 50 g/L was processed with peak area variability of less than 1% for both columns.
Some notes about this work:
• No process development work was done. The first elution peak is slightly under loaded because the first load of
column was based on an assumed titer. A steady state was reached at the second cycle, but with proper
process development, steady state is easily reached at the first cycle.
• In comparing the elution profiles from columns 1 and 2, the differences in the intensity of the two UV signals is
due to a calibration issue.
24© LEWA - Bioprocess Group
Continuous capture delivers higher productivity
with less buffer consumption
Customer A - process development run: 1g/L titer mAb
Triple productivity or 67% less resin and ~ 30% buffer reduction
25© LEWA - Bioprocess Group
Continuous capture delivers higher productivity
with less buffer consumption
Customer A - process development run: 1g/L titer mAb
When comparing the platform batch process to the continuous capture process where the load was
increased from 40 to 50 g/L resin, either productivity was tripled or 67% less resin is used while using
30% less buffer. This is a significant increase in productivity or resin utilization as compared with the
traditional batch process.
26© LEWA - Bioprocess Group
EcoPrime Twin scale-up estimate
Customer A – 1 g/L titer estimate for 100-kg process scale-up
Savings up to $900K in resin or 22 days in processing time.
27© LEWA - Bioprocess Group
EcoPrime Twin scale-up estimate
Customer A – 1 g/L titer estimate for 100-kg process scale-up
This graph demonstrates scale up estimates for both traditional batch and continuous capture
processes that produce 100-kg of purified product based on Customer A’s mAb where operation can
be optimized for either productivity or resin utilization.
Scenario 1 – When using the same amount of resin, the 100-kg of product can be produced in 10
days with a continuous capture process as compared to 32 days with the traditional batch capture
step saving 22 days for processing time.
Scenario 2 – When using the same processing time or throughput (10 days), $900,000 of saving in
resin cost is realized when running the continuous capture process.
28© LEWA - Bioprocess Group
Batch captureLoad 50 g/L resin
Total CV 17.3 L
Continuous capture Load 80 g/L resin
Total CV 1.6 L
Seamless transfer from batch to continuous capture
Customer B – pilot plant process comparison; 5 g/L titer mAb
CVlinear
[cm/h]
Pre‐Sanitization 3 400
Equilibration 3 400
Load 4.6 100
Load interconnected 11.4 150
Wash 1 2 400
Wash 2 5 400
Wash 3 3 400
Elution 3.5 400
CIP 3 400
Re-equilibration 3 400
Sanitization 3 400
Neutralization 2 3 400
Storage 3 400
CVlinear
[cm/h]
Pre‐Sanitization 3 300
Equilibration 3 300
Load 10 150
Wash 1 2 300
Wash 2 5 300
Wash 3 3 300
Elution 5 300
CIP 3 300
Neutralization 1 3 300
Sanitization 3 300
Neutralization 2 3 300
Storage 3 300
Same process steps = simple process transfer
Faster linear velocity = shorter processing time
Process time: 4.5 h Cycle time: 2.5 h
29© LEWA - Bioprocess Group
Batch captureLoad 50 g/L resin
Total CV 17.3 L
Continuous capture Load 80 g/L resin
Total CV 1.6 L
Seamless transfer from batch to continuous capture
Customer B – pilot plant process comparison; 5 g/L titer mAb
Process time: 4.5 h Cycle time: 2.5 h
In the second case study, Customer B used the Contichrom CUBE to obtain initial process
parameters. The process was then scaled-up and optimized on the EcoPrime Twin. To verify the
ability to scale-down, the process from the Twin was re-run on the CUBE.
Shown here is the pilot plant process (5 g/L titer mAb) that was transferred from batch to continuous
capture on the EcoPrime Twin. The process optimization included an increase in load from 50 g/L
resin to 80 g/L resin when going from the batch to the continuous capture process while also
increasing the linear velocity for faster processing.
30© LEWA - Bioprocess Group
Reproducible scale up of 5 g/L mAb capture
EcoPrime Twin run with 14-h process time
peak area peak area
1 0.239 0.276
2 0.255 0.283
3 0.259 0.285
4 0.260 0.286
average 0.2583 0.2849
error %* 1.1 0.7
Column 1 Column 2peak #
Variability of peak
area is ~1 % at
steady state
Contichrom Cube run with 12-h process time
Steady state can be
reached with first
elution.Similar chromatograms when scaling up
31© LEWA - Bioprocess Group
Reproducible scale up of 5 g/L mAb capture
EcoPrime Twin run with 14-h process time
Contichrom Cube run with 12-h process time
Similar chromatograms when scaling up
Shown here is evidence of reproducible scale from the CUBE to the EcoPrime Twin where steady
state continuous operation is reached during the first elution cycle on the Twin. Peak area
reproducibility is ~1% with the pilot scale Twin run.
32© LEWA - Bioprocess Group
Comparison batch to continuous capture
Customer B - process development run: 5 g/L titer mAb
> 2X productivity or 50 % less resin and ~ 50 % buffer reduction
0
5
10
15
20
25
Batch EcoPrime Twin Contichrom Cube
8.4
20.622.2
Pro
ductivity [
g/L
resin
/h]
Productivity - Customer B
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
Batch EcoPrime Twin Contichrom Cube
0.73
0.36 0.34
Buff
er
Co
nsum
ption [
L/g
]
Buffer Consumption - Customer B
33© LEWA - Bioprocess Group
Comparison batch to continuous capture
Customer B - process development run: 5 g/L titer mAb
0
5
10
15
20
25
Batch EcoPrime Twin Contichrom Cube
8.4
20.622.2
Pro
ductivity [
g/L
resin
/h]
Productivity - Customer B
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
Batch EcoPrime Twin Contichrom Cube
0.73
0.36 0.34
Buff
er
Co
nsum
ption [
L/g
]
Buffer Consumption - Customer B
When comparing the platform batch process to the continuous capture process on both the EcoPrime
Twin and CUBE where the load was increased from 50 to 80 g/L resin, either 2x productivity is
realized or 50% less resin is used while using approximately 50% less buffer.
34© LEWA - Bioprocess Group
EcoPrime Twin scale-up estimate
Customer B – 5 g/L titer estimate for 100-kg process scale-up
Savings up to $320K in resin or 6.1 days in processing times
35© LEWA - Bioprocess Group
EcoPrime Twin scale-up estimate
Customer B – 5 g/L titer estimate for 100-kg process scale-up
This graph shows the scale up estimates for both traditional batch and continuous capture processes
that produce 100 kg of purified product based on Customer B’s 5 g/L titer mAb where operation can
be optimized for either productivity or for resin utilization.
Scenario 1 – When using the same amount of resin, the 100-kg of product can be produced in 5 days
with continuous capture as compared to 11.5 days with the traditional batch capture process.
Scenario 2 – When using the same processing time or throughput (5 days), $320,000 of saving in
resin cost is realized when running the continuous capture process.
Contents– Our approach to multi-column capture chromatography
LEWA-Nikkiso America Inc.
36
– Customer case studies
– Conclusions
– A simple set up
– Who we are and what we do
Simple design, simple to set up & run
© LEWA Nikkiso America Inc. 37
Video Link Here
Set up / run system <1 day
Simple design, simple to set up & run
© LEWA Nikkiso America Inc. 38
Video Link Here
Set up / run system <1 day
Because of the simple, straightforward design, the EcoPrime Twin is simple to setup and run. Just
attach to the utilities and columns and you are set to go.
The following is a link to video demonstrating the simple setup. Just copy/paste the link into your
browser.
https://www.youtube.com/watch?v=6v80YZYw7zs&t=5s
Contents
– Our approach to multi-column capture chromatography
LEWA-Nikkiso America Inc.
39
– Customer case studies
– Conclusions
– A simple set up
– Who we are and what we do
Customer experience on mAb shows
Twin column capture delivers:
40
• Significant productivity gains
Better Protein A utilization
• Substantial cost or time savings
Faster loading
Shorter processing time
Reduced buffer consumption
• Seamless scale up
Same process steps
Proven reproducibility
• Simple 2-column approach and system
Accelerate implementation
Minimize risk
© LEWA Nikkiso America Inc.
Customer experience on mAb shows
Twin column capture delivers:
41
• Significant productivity gains
Better Protein A utilization
• Substantial cost or time savings
Faster loading
Shorter processing time
Reduced buffer consumption
• Seamless scale up
Same process steps
Proven reproducibility
• Simple 2-column approach and system
Accelerate implementation
Minimize risk
© LEWA Nikkiso America Inc.
The two customer case studies that were presented demonstrate that for a typical mAb capture step
significant productivity gains and better resin utilization are realized when compared to traditional
batch operation.
Substantial cost or time savings are achieved with faster loading, shorter processing times and
reduced buffer consumption.
Scale up (or down) from the Contichrom CUBE is easy and reproducible because the same process
steps are used with directly scalable column sizes.
All of this is achieved on the EcoPrime Twin LPLC, a simple 2-column system where the less complex
design minimizes downtime, lowers operating cost, and accelerates validation and implementation in
manufacturing.
Contents
– Our approach to multi-column capture chromatography
LEWA-Nikkiso America Inc.
42
– Two customer case studies
– Conclusions
– A simple set up
– Who we are and what we do
LEWA Bioprocess Group
– Engineering
– Fabrication
LEWA-Nikkiso
America, Inc.
– Sales
– Engineering
– Fabrication
LEWA GmbH
Headquarters
Industrial Division
– Sales
– Engineering
– Pump production
– Fabrication
Nikkiso Co., Ltd.
Group
Headquarters
LEWA Bioprocess Group is part of the global family of companies
under the $1.3B US Nikkiso Corporation (1953)
Business sites in >80 countries, >6,000 employees
LEWA Bioprocess Group
– Engineering
– Fabrication
LEWA-Nikkiso
America, Inc.
– Sales
– Engineering
– Fabrication
LEWA GmbH
Headquarters
Industrial Division
– Sales
– Engineering
– Pump production
– Fabrication
Nikkiso Co., Ltd.
Group
Headquarters
LEWA Bioprocess Group is part of the global family of companies
under the $1.3B US Nikkiso Corporation (1953)
Business sites in >80 countries, >6,000 employees
The LEWA Bioprocess group located in Devens, MA is part of the global family of companies of
Nikkiso. With facilities and subsidiaries in 80 countries, we are able to provide support and service for
our products across the globe.
6/21/2017
© LEWA Process Technologies, Inc.45
6/21/201745
GMP systems engineering, manufacturing & automation;• Chromatography• Buffer In-line Dilution• TFF• Downstream process skidsMarkets:• Biopharmaceutical• Pharmaceutical• OEM (bio/pharma)
Devens, MA USA
2000
Acquired in 2014, we are now agroup within LEWA-Nikkiso America, Inc.
The LEWA group ($280M US) is a manufacturer of metering pump headquartered in Leonberg, DE.LEWA was acquired by Nikkiso in 2008. Both Nikkiso & LEWA founded ~65 years ago
LEWA Bioprocess Technologies Group
A group of ~ 60+ professionals dedicated to GMP scale bioprocess systems’ design and manufacture.
Part of the ~ 6,000 employees of Nikkiso Corp. – a diversified company with $1.3B in revenue.
Location
Date of Establishment
Ownership
About LEWA
Video Link Here
Who we are, what we do
6/21/2017
© LEWA Process Technologies, Inc.46
6/21/201746
GMP systems engineering, manufacturing & automation;• Chromatography• Buffer In-line Dilution• TFF• Downstream process skidsMarkets:• Biopharmaceutical• Pharmaceutical• OEM (bio/pharma)
Devens, MA USA
2000
Acquired in 2014, we are now agroup within LEWA-Nikkiso America, Inc.
The LEWA group ($280M US) is a manufacturer of metering pump headquartered in Leonberg, DE.LEWA was acquired by Nikkiso in 2008. Both Nikkiso & LEWA founded ~65 years ago
LEWA Bioprocess Technologies Group
A group of ~ 60+ professionals dedicated to GMP scale bioprocess systems’ design and manufacture.
Part of the ~ 6,000 employees of Nikkiso Corp. – a diversified company with $1.3B in revenue.
Location
Date of Establishment
Ownership
About LEWA
Video Link Here
Who we are, what we do
The LEWA Bioprocess group is a center of excellence dedicated to developing and manufacturing
automated systems for use in GMP scale bioprocessing.
Link to video. Copy/paste the link into your browser.
https://www.youtube.com/watch?v=6IS04Iso-_s&t=39s
A sampling of systems we produce at LEWA Bioprocess
A sampling of systems we produce at LEWA Bioprocess
In addition to process LPLC systems for both traditional and multi-column chromatography, we
manufacture a variety of other systems, including process HPLC, buffer and formulation preparation
systems, TFF, and cell harvest systems all with our control systems and automation software.
LEWA, Creating Fluid Solutions, the LEWA logo, LEWA Intellidrive and LEWA EcoPrime are trademarks of LEWA GmbH.
ChromaCon, Contichrom and CaptureSMB are registered marks of ChromaCon AG.
All goods and services are sold subject to the terms and conditions of sale of the company within LEWA which supplies them. A copy of these terms and conditions is available on request. Contact your local LEWA representative for the most current information.
© 2017 LEWA – Nikkiso America, Inc. – All rights reserved.
For more information: www.lewapt.com
Headquarters LEWA GmbHUlmer Str.1071229 Leonberg GERMANYTel: +49 7152 14-0Fax: +49 7152 [email protected]
LEWA Bioprocess Technologies, Inc. 8 Charlestown RoadDevens, MA 01434 USATel: +978 487 [email protected]
NIKKISO CO. LTD.Yebisu Garden Place Tower 22nd Floor20-3, Ebisu 4-Chome, Shibuya-ku150-6022 TokyoTel: +81 3 3443-3711Fax: +81 3 3473-4963
LEWA-Nikkiso America, Inc.132 Hopping Brook RoadMA 01746 HollistonTel: +1 508 429-7403Fax: +1 508 [email protected]
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