Tx Single Use Seminar Jun09

TangenX - SiusTM Single Use Tangential Flow

Filtration Cassettes

Mark Perreault, TangenX TechnologyMark Perreault, TangenX Technology

TangenX TFF Membranes and CassettesTangenX TFF Membranes and Cassettes

TangenX Technology Corp.– TangenX Technology Corp.– Shrewsbury, MAShrewsbury, MA

• TangenX founded in January 2004 - Shrewsbury, MA

• Novasep acquires a controlling interest in TangenX effective September 1, 2006

• TangenX and Novasep introduce validated TFF reusable products at IBC in Boston, October 2007

• TangenX and Novasep introduce industry’s first validated TFF reusable products at IBC in California, Dec. 2008

• + 40 years Membrane and TFF Experience

TangenX Overview Ultrafiltration Membranes

TangenX Membranes - mPESTangenX Membranes - mPES



“Single Cast”, uniquely controllable process– Reduced number of manufacturing steps

• Lower cost and excellent consistency and reliability– Tightly controlled microporous/UF

transition interface. • Balanced flux and selectivity• No breakthrough of the UF skin• Macroporous substructure

for maximized flux

SEM X-Section of the ProStream 100KD

Integral “cast-modification” of the membrane surface chemistry–Addition of a second (hydrophilic) polymer into the pre-casting membrane solution–Consistent surface modification that deliver

•Lower protein binding•Excellent chemical resistance


TangenX mPES MembranesTangenX mPES Membranes

Two (2) Membrane ChemistriesTwo (2) Membrane Chemistries

Modified Polyethersulfone (PES) membranes:Modified Polyethersulfone (PES) membranes:• ProStreamProStreamTMTM: : 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, 100kD, 300kD, 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, 100kD, 300kD, 0.1µm, 0.2µm0.1µm, 0.2µm–Neutrally Balanced Charge Neutrally Balanced Charge –Low Protein BindingLow Protein Binding

•HyStream HyStream TMTM: : 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, 100kD, 300kD, 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, 100kD, 300kD, 0.1µm, 0.2µm0.1µm, 0.2µm

–Hydrophilic - Minimal fouling with hydrophobic molecules Hydrophilic - Minimal fouling with hydrophobic molecules –Low Protein BindingLow Protein Binding

ProStreamTM 10 kD (1000X)

HyStreamTM 10 kD (1000X)

TangenX offers two membrane chemistries; ProStream and TangenX offers two membrane chemistries; ProStream and HyStreamHyStream Available in multiple cutoff’s: 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, Available in multiple cutoff’s: 1kD, 3kD, 5kD, 10kD, 30kD, 50kD, 100kD, 300kD, 0.1µm, 0.2µm100kD, 300kD, 0.1µm, 0.2µm

ProStream

• mPES membrane cast on a polypropylene non-woven• The membrane is neutrally charged, rendering it low binding for enhanced flux and recovery• The membrane has excellent chemical resistance and very low extractables.

HyStream• mPES membrane also cast on a polypropylene non-woven• It’s modification renders it extremely hydrophilic in nature, resisting fouling from hydrophobic species such as fat, lipids, and various antifoam agents.• The membrane has good chemical resistance and offers an alternative to RC membranes in some cases.

ProStream 10KD HyStream 10KD

(+) (-) (+) (-) (+) (-) (+) (-) (+) (-) (+) (-) (+) (-) (+) (-)

ProStream

(+/-)

(+/-)(+/-)(+/-)

(+/-)

(-) (-) (-) (-) (-) (-) (-) (-) (-) (-) (-) (-) (-) (-) (-)

Un-Modified PES (+)(+) (+)(+) (+)(+)




• Membranes were challenged with model proteins and the protein rejection was quantified

• The rejection of each protein for a given MWCO membrane was plotted against the protein’s molecular weight

• The selectivity curves generated show discrete selectivity profiles with no overlap

• TangenX membranes demonstrate exceptional selectivity when challenged with “model” proteins in solution

Membrane Selectivity Curves for:

ProStream Membranes

HyStream Membranes

Contact Angle Measurement

(water at 20oC)

HyStream

Molecular Weight vs. % Rejection

0102030405060708090

100

0.1 1 10 100 1000 10000

Molecular Weight (kD)

% R

ejec

tio

n

1K

3K

5K

10K

30K

50K

100K

300K

0.1um

SEM of the HyStream 0.1um surface at 20,000X

Data generated in triplicate using 13cm2 stirred cells



• Typical 3 to 6 rule for MWCO selectionTypical 3 to 6 rule for MWCO selection

• Better selectivity allows users to use a higher MWCO membrane for improved flux. • TangenX membranes can completely retain molecules as little as 2X different in MW.

• Example: •TangenX 100KD ProStream modified PES

• Purified MAb concentration, 10mg/ml - (~150KD)

• Typical flux is 90 to 100 LMH

• Greater than 99% retention


100KD ProStream mPES 100KD ProStream mPES MAb Optimization – 10mg/mlMAb Optimization – 10mg/ml


• Typical 3 to 6 rule for MWCO selectionTypical 3 to 6 rule for MWCO selection

• When fractionating, better selectivity allows users to separate molecules closer in molecular weight. • TangenX membranes can fractionate molecules as little as 2X different in MW.

• Example: •TangenX 100KD ProStream modified PES

• Purified BSA concentration, 1.0mg/ml - (67KD)

• Typically less than10% retention means BSA is not retained by the 100KD membrane


TangenX membranes offer greater flux & selectivity than TangenX membranes offer greater flux & selectivity than traditional mPEStraditional mPES


TangenX - SiusTM Single Use Tangential Flow

Filtration Cassettes

ISPE Carolina South-Atlantic Chapter 25

• Single-use technology has made bioprocesses:• Cleaner• Safer• More Efficient

• Single-use technology has enabled a typical purification process to eliminate numerous costly clean in place (CIP) operations

SiusSiusTMTM Single-Use TFF Cassettes Single-Use TFF Cassettes

ISPE Carolina South-Atlantic Chapter 26




• The first truly disposable TFF cassette designThe first truly disposable TFF cassette design

• The SiusThe SiusTMTM cassette incorporates a unique patented* cassette incorporates a unique patented* design with a more automated process for assembly.design with a more automated process for assembly.

• Cassettes are direct retrofits for all existing TFF devicesCassettes are direct retrofits for all existing TFF devices

*patent pending


• Full range of formats and MWCO (molecular weight cut offs)

• Pre-sanitized with 0.2M Sodium Hydroxide • Certified low TOC and endotoxin

• Equilibrate & ready-to-use

• Economic TFF Solution

Benifits – SiusBenifits – SiusTM TM CassettesCassettes

• SiusSiusTMTM-LS – from laboratory to small pilot scale-LS – from laboratory to small pilot scale

• Available in 0.1mAvailable in 0.1m22, 0.2m, 0.2m22, and 1.0m, and 1.0m22

• Stack cassettes up to 0.5mStack cassettes up to 0.5m22

• SiusSiusTMTM – from pilot to process scale – from pilot to process scale

• Available in 0.5mAvailable in 0.5m22, 1.5m, 1.5m22, and 2.5m, and 2.5m22

• Stack cassettes up to 7.5mStack cassettes up to 7.5m22

Linear Scalability – SiusLinear Scalability – SiusTM TM CassettesCassettes

“L” Screen

1.0mm Open Channel

Flow Spacers

Low Viscosity High Viscosity & Particulates

0.5mm Open Channel

Membrane

Flow Spacers

Lower Cross Flow Rate

Higher Cross Flow Rate

Higher Cross Flow Velocity Lower Cross Flow Velocity

Channel Options – SiusChannel Options – SiusTM TM CassettesCassettes

• Sius™ cassettes have been designed and constructed using FDA approved materials that have been validated for use for demanding biopharmaceutical applications.

•The Sius™-LS laboratory scale and Sius™ process scale TFF cassettes are purpose-built for single use processing with optimal performance that is equivalent to our NovaSet reusable product line. •Each cassette is manufactured in a fully validated and documented manufacturing process according to the principles of cGMP and meet specified release criteria.

•Cassettes are manufactured using approved standard operating procedures and individually tested for both air integrity and hydrodynamic performance.

Manufacturing – SiusManufacturing – SiusTM TM CassettesCassettes

• SiusSiusTMTM Single Use TFF Cassettes utilize an innovative 2 stage assembly process

• Membranes and permeate channels are bonded together

Manufacturing – SiusManufacturing – SiusTM TM CassettesCassettes

• Retentate channels consist of rigid plastic frames and turbulence promoting screens

• The assembly is instantly bonded together using medical grade silicone pressure sensitive adhesive

• Benefits• Process mfg. efficiency greatly

improved• Reduced costs• Reduces lot rejection

Patent Pending

• Materials of ConstructionMaterials of Construction– MembraneMembrane PESPES– Membrane SupportMembrane Support PolypropylenePolypropylene– ScreenScreen PolypropylenePolypropylene– Channel SpacerChannel Spacer HDPEHDPE– Filtrate SealsFiltrate Seals UrethaneUrethane– GasketGasket EPDMEPDM

• SpecificationsSpecifications– Maximum PressureMaximum Pressure

• 7 bar (100 psi) forward; 0.33 bar (3 psi) reverse7 bar (100 psi) forward; 0.33 bar (3 psi) reverse– Air IntegrityAir Integrity

• UF cassetteUF cassette ≤ 30 sccm @ 1 bar (15 psi)≤ 30 sccm @ 1 bar (15 psi)• MF cassetteMF cassette ≤ 30 sccm @ 0.2 bar (3 psi)≤ 30 sccm @ 0.2 bar (3 psi)

– Recirculation FlowRecirculation Flow• 'L' Screen'L' Screen 6-9 lpm/m2 @ 0.7 bar (10 psi)6-9 lpm/m2 @ 0.7 bar (10 psi)• 'J' Channel'J' Channel 1-1.5 lpm /m2 @ 0.3 bar (1.5 psi)1-1.5 lpm /m2 @ 0.3 bar (1.5 psi)

• ManufacturingManufacturing– Fully validated process, developed and documented in accordance with cGMP principlesFully validated process, developed and documented in accordance with cGMP principles– Quality Control lot release testing includes 100% device integrity and hydrodynamic testingQuality Control lot release testing includes 100% device integrity and hydrodynamic testing– Sanitized and packaged in 0.2M NaOHSanitized and packaged in 0.2M NaOH– Manufacturing process is fully validated, a validation guide is available to referenceManufacturing process is fully validated, a validation guide is available to reference

Specifications – SiusSpecifications – SiusTM TM CassettesCassettes

Product Support – Validation GuideProduct Support – Validation Guide

Product Support – C of CProduct Support – C of C

Product Support – Product UseProduct Support – Product Use

Product Support – Product UseProduct Support – Product Use

• Sius™ cassettes have been designed and constructed using FDA approved materials that have been validated for use for demanding biopharmaceutical applications.

•The Sius™-LS laboratory scale and Sius™ process scale TFF cassettes are purpose-built for single use processing with optimal performance that is equivalent to our NovaSet reusable product line.• •Each cassette is manufactured in a fully validated and documented manufacturing process according to the principles of cGMP and meet specified release criteria.

• Internally audited by Novasep QA

•Cassettes are manufactured using approved standard operating procedures and individually tested for both air integrity and hydrodynamic performance.

Product Support – ManufacturingProduct Support – Manufacturing

41

• Single-Use TFF eliminates the need to flush with costly purified water or measure water permeability rates, saving time and resources.

• Pre-Sanitized cassettes are installed, equilibrated with buffer, and used in processing.

• Development work, scale up and processing can be conducted nearly 50% faster without the need for membrane cleaning and measurement or water flux recovery.

• Cassette performance is more consistent from run to run as each process occurs using a new membrane.

• Cross-contamination is minimized with single-use cassettes.

Single Use - SummarySingle Use - Summary

TangenX - NovaSetTM Tangential Flow Filtration

Cassettes

NovaSetNovaSetTMTM-LS Reusable TFF Cassette-LS Reusable TFF Cassette

NovaSetNovaSetTM TM Reusable TFF CassetteReusable TFF Cassette



NovaSetNovaSetTMTM- Feed Channel Options- Feed Channel Options

“H” Screen

Membrane

Suspended Screen

Flow Spacers

“L” Screen

Low Viscosity High Viscosity & Particulates

Higher Cross Flow Velocity Lower Cross Flow Velocity

Lower Cross Flow Rate

Higher Cross Flow Rate




NovaSet™ Cassette0.5, 1.5 or 2.5m2

• Specifications– Maximum Pressure

• 7 bar (100 psi) forward; 0.33 bar (3 psi) reverse

– Recirculation Flow• 'H' Screen 6-9 lpm @ 1

bar (15 psi)• 'L' Screen 6-9 lpm @

0.7 bar (10 psi)• 'S' Channel 1-1.5 lpm @

0.3 bar (1.5 psi)• Manufacturing

– Fully validated process, developed and documented in accordance with cGMP principles

– Quality Control lot release testing includes 100% device integrity and hydrodynamic testing

– Individually packaged in a sealed foil lined bag and outer cardboard carton

– Wet packed in 20% Glycerin and 0.1% Sodium Azide

•Materials of Construction–Membrane: PES–Membrane Support: Polypropylene–Screen: Polypropylene–Encapsulant: Urethane–Gasket: Platinum Cured Silicone

• Sius and NovaSet cassettes provide an interchangeable TFF platform

• Same membrane used in both products• Comparable cassette performance• TangenX cassettes allow scale up and scale

down between both cassette formats

NovaSetNovaSetTM TM vs. Siusvs. SiusTMTM TFF Cassette PerformanceTFF Cassette Performance


• Pressure drop vs. crossflow rate confirms scalability within the same product family

• Also confirms interchangeability between family of cassettes

• TangenX cassettes allow scale up and scale down between both cassette formats

5X Concentration of BSANovaSet-LS vs. Sius-LS

0

20

40

60

80

100

120

140

160

180

200

Filtrate Flux (LMH) % Recovery

NovaSet LS

Sius LS


• 5 mg/ml BSA• TMP 25psi• CFF = 0.6L/min/ft2

• Flux and recovery comparable• TangenX cassettes allow scale up

and scale down between both cassette formats

• Application: 5X BSA concentration

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TangenX - TFF Cassette Holdersfor

NovaSetTM and SiusTM

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BioPharm TFF – LaboratoryBioPharm TFF – Laboratory

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BioPharm TFF – PilotBioPharm TFF – Pilot

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BioPharm TFF – ProcessBioPharm TFF – Process

• NovaSet-75 Hydraulic TFF Cassette Holder

• NovaSet-300 Hydraulic TFF Cassette Holder

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TangenX - SiusTM Filter Plate Insert (FPI)

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• The FPI follows the disposable The FPI follows the disposable TFF cassette designTFF cassette design

• FPI direct retrofits for all existing FPI direct retrofits for all existing TFF devicesTFF devices

• The filter plate insert system The filter plate insert system provides a disposable flow path for provides a disposable flow path for Sius cassettesSius cassettes

Disposable Flow Path forDisposable Flow Path for SiusSiusTM TM CassettesCassettes

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Filter Plate Insert (FPI)

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System Design

• Insert plates may be used with existing filter holders by replacing cassettes

• All threaded connections in the fluid path are isolated by an o-ring – "sanitary design“

• Low cost dedicated holder available

System Design

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• Type 304L/316 Stainless Steel– Clamp Base, Cover– Washers / Spacers– Screws

• Brass Nuts– 1-¼" Socket Hex Nut

• Polyethyelene (Contact Surface)– Filter Plate Insert– Barbed Fittings

• Polypropyelene (Contact Surface)– Luer Lock Fittings

• EPDM (Contact Surface)– O-Rings

Materials of Construction

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Insert Plate Features

• Each plate has (1) set of Feed/Retentate Ports and (1) set of Filtrate Ports

• Each plate has (2) male luer lock pressure tap connections (supplied with caps)

• Plates are approximately 1" thick with each port entrance clearly identified

• Each plate is sanitized, dried and individually bagged

– Includes CofC/A for traceability

• Compatible with NovaSet reusable cassettes.

Insert Plate Features

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• Similar design and function to process plates

• To be used in NovaSet-LS hardware

FPI - Laboratory Scale

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Options

• Mini T/C or 1" T/C Fitting replaces Barb (not shown)

• Analog Gauge Option

FPI - Options

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• The filter plate inserts (FPI) are available The filter plate inserts (FPI) are available for Beta testing – June 09for Beta testing – June 09

• The functionality of the FPI with The functionality of the FPI with competitor's holders is underway competitor's holders is underway

• Various hose barb and instrument Various hose barb and instrument connection ports are available, no connection ports are available, no standard has been established yetstandard has been established yet

• Filter plate inserts will be provided with:Filter plate inserts will be provided with:• MSDS (material safety data sheet) • COA (certificate of analysis) • COO (certificate of origin) BSE statement • shelf life• Extractables & leachables

Current Status of FPICurrent Status of FPI

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TangenX Overview

Thank You For Your Attention

Documents

Tx Single Use Seminar Jun09