Advanced Degrees of Separation

Latest Advances
in
Tangential Flow Filtration
for
Separation and Concentration
 The NCSRT Challenge

Develop novel filtration, separation and

purification technologies to elevate performance
optimization standards in bioprocessing,
development and production.
 The NCSRT Focus
Create innovative filtration modules that:
• improve fluid dynamics
• optimize membrane use
• expand performance limits…

…well beyond those produced using traditional

Tangential Flow Filtration (TFF)
 The NCSRT Solution
Apply a revolutionary NCSRT design enabling bioprocessing
professionals to increase optimum yield, press for greater standards
of purity and achieve:
• Higher Permeate Rates
• Improved Cleaning
• Increased Process Yield
• Higher Cell Densities
• Finer Separations
• Higher Protein Concentrations
• Higher Viscosities
• And Consistent, Repeatable Results
 The NCSRT Advantage
In Downstream Processing

Typical Cell Culture Flow Diagram

 The NCSRT Solution:
OPTISEP® SmartFlow™

The Technology
Proven
To Heighten
TFF Processing Yield
By 50% To 500%

Tangential Flow Filtration (SF-

(SF-TFF)
 Separations of Significance

NCSRT Application # 129:

Protein Concentration / Diafiltration

Concentration Step 0.4 to 4% Solids Average

Average (LMH)

™ ™
- Omega 10 kD Maximate 30
- GE Amersham 10 kD Hollow Fiber 39
™
- NCSRT OPTISEP RC 10 kDa 70
Customer Commentary:

“The NCSRT module performs best compared with any other formats used.”

“Flux was stable at 70 LMH through two hours of concentration and 3X diafiltration.”

Omega™ and Maximate™ are registered trademarks of Pall Corporation. OPTISEP™ is a registered trademark of NCSRT, Inc.
Separations of Significance

NCSRT Application # 272:

Fungal Culture Processing
Avg. (LMH)
• 750 kD GE Amersham 35
• OPTISEP® PES 1000 kD 66
• OPTISEP® PES 500 kD 180

Concentration Step
• 10 kD GE Amersham 10
• OPTISEP® PES 10 kD 60

Comparative Data Summary:

Product Yield for GE Amersham 26%
Product Yield OPTISEP 96%
Separations of Significance
NCSRT Application #104:
Protein Percipitates
(LMH) Avg. Concentration
Step 0.4 to 4% Solids
• 0.2 PVDF Millipore Cassette 40-
40-5 22
• 0.2 PVDF Millipore ProStak 30-
30-0 0
• 0.1 PVDF NCSRT OPTISEP 350-
350-150 250
• RC 100 kD NCSRT OPTISEP 200--175
200 187.5

Diafiltration Step 10X

• 0.2 PVDF Millipore Cassette 5 5
• RC 100 kD NCSRT OPTISEP 186 186

Final Process:
RC 100 kD NCSRT OPTISEP – Concentration from 0.4 to 20% Solids
and 10X Diafiltration with an average flux rate of 186 LMH
Separations of Significance
NCSRT Application #79:
E. coli Lysate Processing
Filter Type Avg. (LMH)

• Pellicon ™ Zero
• Centrasette ™ Zero
• 3 mm Hollow Fiber 5

• OPTISEP® PS 0.2 18
• OptiSEP® PES 1000 kD 30
• OptiSEP® PES 500 kD 70

Comparative Data Summary:

Yield from Hollow Fiber 50%

Yield from OPTISEP 94%

Pellicon™ is a registered trademark of Millipore, Inc. Centrasette™ is a registered trademark of Pall Corporation.
Separations of Significance

NCSRT Application #133:

Membrane Stability

Processing Performance Summary

• Membrane delivered to bioprocessing customer (1999)
• 5 years old, 0.875 mm channel, 2.5 kD, 5 square meters
• Processed monthly batches for 2 years
• During 2002 – added second 5 square meter module
• Increased process duration to 8 days (24 hours per day)
• Membrane cleaned / turned around in 12 hours
• Process restarted immediately without change in flux or retention
 Key Findings:

NCSRT, Inc. research verifies that fluid distribution is the

the
single most important factor impacting:
• flux decay
• cleaning validation
• and scale up

Field Data Analysis Summary :

Uniform fluid distribution is required for true optimization

of TFF processes.
 Optimal Flow: Hollow Fiber

Identify
the
path
of
least
resistance
 Flow Geometry: Hollow Fiber

• Fibers in center are

blasted by turbulence

• Fibers along the

outer diameter are
stagnant

• Red = Optimal Flow (path of least resistance)

• Purple = Secondary Flow
• Blue = Decreasing Flow
• Green = Poor Flow
• Orange / Yellow=
Yellow= Poorest Flow
 Flow Dynamics: Conventional Filter
Red = Optimal Flow
(path of least resistance)
Purple = Secondary Flow
Blue = Decreasing Flow
Green = Poor Flow
Orange / Yellow=
Yellow= Poorest Flow
 Retentate Channels: Conventional Filter
How do you validate cleaning
the “X” areas
between the retentate channels?

Red = Optimal Flow

(path of least resistance)
Purple = Secondary Flow
Blue = Decreasing Flow
Green = Poor Flow
Orange / Yellow=
Yellow= Poorest Flow
Flow Distribution: Conventional Filter

INLET OUTLET
 The OPTISEP Advantage

SmartFlow™ TFF …Optimizing Fluid Dynamics

SmartFlow™
 OPTISEP SmartFlow TFF Models

• NCSRT’s patented rib design creates uniform retentate channels

• Produces equal flow rates across the entire membrane surface.

• To optimize for viscosity and solids content, ribs are availabl

available
e
in multiple channel heights
Note the clear advantage SmartFlow technology provides
by equalizing flow across 100% of the membrane surface
 NCSRT Retentate Channels

These patented retentate channels, found only in NCSRT filters

also reduce gel layer buildup and prevent fouling
 OPTISEP Modules
Exponentially Advance TFF Performance

Bioprocessing professionals throughout the world continue to

document how the entire line of OPTISEP modules consistently
provide:

• Protein Recoveries >99%

• Cell Mass Concentrates to >80% Solids (wet weight)
• and Protein Concentration to >40% Solids
 OPTISEP ® 800
• OptiSEP 800 is ideal for
membrane and process
1--20L
evaluation from 1

• Unique flow channels utilize 100%

of membrane surface

• Autoclavable and can withstand

multiple SIP cycles

• Available with channel heights of:

! 0.25 mm
! 0.50 mm
! 1.0 mm
! 1.5 mm
 OPTISEP ® 3000
• OptiSEP 3000 series filter modules yield
optimum efficiency on a range of
solutions from … dilute
proteins to viscous cell harvesting

• Homogeneous fluid dispersion

• Available in wide range of membrane

materials and pore sizes

• Ideally suited for downstream processing of 2-

2-
50 L bioreactors
 OPTISEP ® 7000
• Homogeneous fluid distribution

• Yields optimum efficiency on a range of

solutions from…
dilute proteins to viscous cell
harvesting

• Multiple channel heights formed

by NCSRT’s patented distribution ribs

• Efficiently utilizes 100% of

the membrane area
 OPTISEP ® 11000
• OptiSEP 11000 series filter modules yield
optimum efficiency on a range of solutions
from… dilute proteins to viscous cell
harvesting

• Available in wide range of membrane

materials and pore sizes

• Production systems processing

50 – 1,400,000 L per day in operation
 PUROSEP™ LT-1
• Designed for dependable laboratory
scale filtration
• Evaluate and develop micro and
ultra filtration separation processes
• Portable
• Countertop system

Shown with
OPTISEP®-200
filter module
 PUROSEP LT-2
Dependable laboratory
scale filtration
Evaluate and develop
micro and ultra
filtration separation
processes
Performance levels of
larger units
Compact
Countertop System
Compatible with
OPTISEP® 200 -
3000 filter modules
 PUROSEP LT-2
Provides dependable laboratory
scale filtration to evaluate
micro and ultra filtration
separation processes.
Offers performance levels of
larger units.
Precise control of recirculation
flow rate, permeate flow rate
and transmembrane pressure
ensures that test data can be
directly scaled for production.
Shown
Shownwith
Allows easy evaluation of with
optional
optionalcart
different filter formats, cart
membrane types and
porosities.
 PUROSEP PT
Built-in process tank for totally self-
contained operation.
GMP compliant for high value, low
volume feed stocks for clinical
production.
Able to handle 100-fold
scale up.
Designed for filtration and
concentration of 10 to 1,000 L of
feed stock to less than
1 L of product.
Allows easy evaluation of different
filter formats, membrane type
and porosities.
 PUROSEP PRO
Designed for a wide range of
tangential flow microfiltration
and ultrafiltration
applications.
Utilized worldwide for
concentration or purification
of cells, proteins, blood
fractions, diafiltration, and
many other processes in the
pharmaceutical and
bioprocess industry.
SIP and CIP compatible.
Designed for large scale
production using a feed and
bleed loop.
Operates with a variety of
membranes, filters and
process streams.
Chromatography System
SYSTEM FEATURES
"Bubble Trap

"Column Control Valve

"CIP and SIP Compatible

"UV

"pH

"Conductivity

"Temperature

"Pressure

"Precolumn Filtration

"0-100% Gradient by
Metering Pumps
"Isocratic
 NCSRT Solution
• Validation Services
• Process Development
• Process Optimization
• Training Seminars
• Preventive Maintenance
 Thank You
• Rental and Demonstration Units
Available

NCSRTinformation@ncsrt.com

919-387-8460