NuSep Presents at Bioplasma Conference2013/09/06 · Prakash Patel Managing Director +61 2 8415 7300 [email protected] About NuSep NuSep (ASX: NSP) is a publicly listed life

NuSep Holdings Ltd 30 Richmond Rd

Homebush NSW 2140

Postal Address P.O. Box 2202

Homebush West NSW 2140

Contact Details

Telephone +61 2 8415 7300

Facsimile +61 2 8415 7399

Email [email protected]

Web www.nusep.com

ABN 33 120 047 556

FOR IMMEDIATE RELEASE

NuSep Presents at Bioplasma Conference

Sydney, Australia 6th September 2013–NuSep Holdings Ltd (ASX:NSP) wishes to advise the market that Dr Hari Nair, Executive Chairman of PrIME Biologics Singapore presented one of the key note addresses at the Bioplasma World Asia 2013 conference held on 4th and 5th of September. This presentation was on the PrIME mini mill plasma fractionation concept and the roll out of the first a plasma fractionation facility in South East Asia. Dr Nair also participated in a number of panel discussions. The conference was well attended including members from the global fractionators and regulators, both European and Asian. For more information on this conference see http://www.imapac.com/index.php?page=BioplasmaWorldAsia2013. Attached is a copy of the presentation. - END- For more information please contact: Contact: Prakash Patel Managing Director +61 2 8415 7300 [email protected] About NuSep NuSep (ASX: NSP) is a publicly listed life sciences company that sells products into the global BioSeparations market. The company has offices in both Sydney Australia and Atlanta, USA. With a 30 year heritage in biological separations, NuSep has forged a world class reputation for its innovative yet simple biological separation techniques based on its PrIME1 technology. The PrIME technology has produced a number of world firsts including the world’s first IVF sperm separation device. In short NuSep has redefined the BioSeparations market through innovation and simplification. NuSep’s world renowned research team has developed an extensive portfolio of patented products. In all, NuSep currently manufactures, distributes and sells 55 products to customers in the USA, Europe, Asia and Australia. NuSep Products:

Gels – NuSep manufactures and sells precast gels including the innovative nUView Gels, which can be visualised 2 minutes after use.

1 PrIME stands for Preparative Isolation by Membrane Electrophoresis

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Separation Instruments – NuSep has developed two unique biological separation instruments. The ProteomeSep can separate biological samples into 8 fractions for use in the proteomic market. The SpermSep separates sperm for fertility treatments such as IVF.

Biological Products – NuSep supplies research grade biological products manufactured using its unique separation technologies. These products include human IgG and Albumin.

For more information about NuSep please visit the company’s website www.NuSep.com About PrIME Biologics Pte Ltd PrIME Biologics is a Singapore based biotech company that has developed a disposable therapeutic plasma manufacturing process based on NuSep’s PrIME Technology. PrIME Biologics process is based on the Preparative Isolation by Membrane Electrophoresis (PrIME) technology developed by NuSep. PrIME provides disposable modular processing that is ‘electronically’ driven membrane fractionation. The PrIME process increases product yields to over 90% relative to the existing process 50% while increasing product safety. The PrIME has been shown to remove virus, bacteria, endotoxins and prions which cause ‘Mad Cow Disease’. Further, PrIME Biologics process can produce multiple plasma products in hours compared to days, which is required by the current manufacturers. The initial application for the PrIME process will be the Currently Unprocessable Plasma which represents approximately 50% of all the plasma collected in many Asian countries.

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Dr Hari Nair

A Solution to Developing Plasma Fractionation for Asia: A PrIME objective

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Blood product market overview

•US$15 billion market worldwide. Dominated by 4 manufacturers.•23‐25M L of plasma fractionated globally – 35% recovered plasma*•Estimated 5.8M litres are destroyed in the developing word*•PrIME to address existing and open untapped multi US$B market by:

►Using the PrIME to process Plasma currently thrown away using a:►Disposable manufacturing process that can►Process any volume from 20L ►Double the volume of finished product while increasing safety

Plasma fractionation in the Asian region constitutes only 20% of the total volume fractionated worldwide for a region that houses 60% of the world population

*A.M. Cheroghali. H. Abolghasemi Biologicals 38 (20 10) 81 -86

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World Supply of Plasma

North America5% of the worlds population18.2m L of plasma collected

Europe10% of the worlds population16.6m L of plasma collected

References1.Worldwide Demand/Supply of Plasma and Plasma-derived medicines, Patrick Roberts, MRB 10 May 20112.Improving availability and affordability of plasma-derived medicines, Abdol Majid Cheraghali, Biologicals 38 (2010) 81- 86

Rest of the World Rest of the World 5.8m L of plasma discardedF

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World Market for Plasma Products

Asia Asia 60% of the worlds population60% of the worlds population1515--20% of the world20% of the world’’s plasma productss plasma products

North America5% of world population35% of the world’s plasma products

Europe10% of the worlds population35% of the worlds plasma products

References1.Worldwide Demand/Supply of Plasma and Plasma-derived medicines, Patrick Roberts, MRB 10 May 20112.Plasma fractionation in Asia–Pacific: challenges and perspectives, T. Burnouf, ISBT Science Series (2011) 6, 366–372

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The Asian Market Opportunity is significant because:•Requirement for self‐sufficiency•There is excess demand;•Current processes do not meet the Asian requirement;•Blood and plasma are emotive issues;•Supply is Country specific and centrally controlled although stored separately;•Demand is inelastic because of the limited supply.

Asian Market Opportunity

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“Production of sufficient quantity of qualified plasma is the key element in the availability of plasma derived medicines”

•Enforcement of FDA level plasma collection processes in third world countries can be prohibitive. •The challenge is to introduce a technology which can address concerns about product safety thus enabling the processing of the large volumes of discarded plasma whilst improving quality of plasma collected

Asia needs a new approach

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• Electrophoresis (Electrical Charge) based separation

• Buffer determines pH• Separation using polyacrylamide membranes with nominal molecular weight cut‐off (MWCO)

PrIME Technology(Protein Isolation by Membrane Electrophoresis)

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Membrane Interior cross section‐ x400

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Restriction Membrane


Separation Membrane

Stream 2

Stream 1

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Stream 2

Stream 1



Separation Membrane

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Stream 2

Stream 1



Separation Membrane

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Sample Stream


Restriction MembraneF

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• High protein recovery (generally over 80%)• High Resolution of separation• Faster separation• Easily scalable • Separations by size and/or charge• Proteins remain in liquid medium• Carrier ampholytes and isoelectric membranes not required

for the separation• Purification without pressure

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►Improved product safety through• Viral reduction• Prion reduction• Pyrogen reduction• S/D and PrIME integration

►No contact between plasma and production equipment eliminates CIP►Can be Complimentary to Current technology►Compatibility with chromatography polishing

Added Benefits

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Membrane size scale up

Membrane SurfaceArea (cm2)

Scale Up

MF10 1.5

BF100 15 100X

GF100 5X31.5X31.5 3310X

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PrIME Process development for:– IgG‐ IVIG and hyperimmune– Albumin– Factor VIII– Alpha‐1‐proteinase inhibitor– Antithrombin III– Fibrinogen

Product safety validation for:– Viral clearance (enveloped and non‐enveloped viruses)– TSE/Prion clearance– Endotoxin removal

Blood product development and safety validation

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pH 5.4 bufferpH 5.4 buffer

pH 5.4 bufferpH 5.4 buffer

STREAM 1STREAM 1

STREAM 2STREAM 2‐

+ + ‐‐‐‐

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1X

100X

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Retention Time (min)

Therapeutic IVIG PrIME IgG

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IgG1 IgG2 IgG3 IgG4

010203040506070

% T

otal

IgG

Pooled Plasma

8-1-01 GL IgG pool

9-1-01 GL IgG pool

IVIG

IgG1 IgG2 IgG3 IgG4

Pooled Plasma 65 23 3.8 4.2PrIME IgG 63 24 3.1 <4.3PrIME IgG 67 27 3.1 <5.0IVIG 67 22 3.0 3.0

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IgG IgA IgM Transferrin Albumin

PrIME IgGS1end Phase 1Starting plasma

0%

20%

40%

60%

80%

100%

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PrIME IgG

S1end Phase 1

Starting plasma

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Assay Target Specification Averaged Results from PrIME process

1D SDS PAGE ≥ 95% 100 %

IgG Aggregation ≤ 2% IgG Aggregation 0.28 %

Total Protein For Purity Assessment 49mg/mL

Total IgG ≥ 90% Recovery from pooled plasma 91%

IgG Subclasses Ratio corresponds to pooled plasma Corresponds

IgA No specification 0.12mg/mL 5% solution

IgM No specification <0.07mg/mL

Plasminogen/Plasmin No specification 7.3 CTAu/mL 5% solution

pH No specification 4.92 (formulated)

Osmolality No specification 630 (formulated)

Residual Acrylamide Internal Specifications BLD

Endotoxin < 5 EU/mL

Anti‐D hyperimmune product specifications

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Flow Flow

B‐19

B‐19

B‐19

B‐19

B‐19

IVIG

IVIG

IVIGIVIG

IVIG

IVIG

IVIG

IVIG

Pressure forces Viruses through Nano-Filtration filters causing contamination

Viral Clearance: Current Technology Using Nano Filtration

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B‐19

B‐19

B‐19

B‐19B‐19

IVIG

IVIG

IVIGIVIG

IVIG

IVIG

IVIG

IVIG

+ + + + + + + + + +

‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐

In the PrIME Viruses are negatively charged and stay in the positive chamber

Viral Clearance: Using PrIME

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Virus Model virus

Family Genus Genome Envelope Size (nm)

B19 PPV/CPV Parvo Parvo ssDNA no 18-24HCV BVDV Flavi Pesti ssRNA yes 40-60HIV BVDV Flavi Pesti ssRNA yes 40-60HAV - Picorna Hepato ssRNA no 22-30HBV DHBV Hepadna Avihepadna DNA yes 40-48

Model viruses for blood borne pathogens

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Evaluation of Viral Clearance by PrIME process

– Scale down laboratory model of manufacturing process

– Model viruses selection– Spike with high titer stocks of virus

• Log clearance is dependent on starting virus stock

– Quantify reduction of virus• Reproducible, sensitive assay methodology (infectivity/PCR or RT‐PCR)

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GelVirus

PET Fibres

Up Stream

VIRUS RETENTION 300 kD Membrane, t=30 mins, x70,000

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PPV removal during IgG separation by PrIME process

SampleNested PCR (log10 GE*)

Infectivity (log10TCID50)

S1 0 hr 7 7.5

S1 residual 6 6.4

IgG product 1 <1.9

Viral reduction > 6 > 5.6

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TSE removal during IgG separation by PrIME process

Buffer conditions

Starting sample

Product sample

(IgG)

Reduction Factor (log10)

IgG separation

1.3 x 104 7.0 x 100 ≥ 3.3

S10 S1end S2end

1 2 3 4 5 6 7 8 910 1 2 3 4 5 6 7 8 910 1 2 3 4 5 6 7 8 910

Microsomal PrPSc

1 2 3 4 5 6 7 8 910

1 – Straight positive 10-2.1

2 – MW marker3 – Undigested neat 4 – Digested neat5 – Digested 10-0.7

6 – Digested 10-1.4





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PrIMEProduct

TherapeuticIVIG

Source material plasma plasma

Purity > 99 % > 99 %

Recovery >85 % ~40%

PPV removal > 4 log > 8 log

Endotoxin removal >99 % >99 %

Prion removal BLD* ?

**BLD – Below Limit of Detection** 8 log removal is achieved by two viral clearance steps, each of which demonstrated greater than 4 log removal

PrIME Technology v’s Current Available Therapeutic Products

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Selectivity

Process Speed

CapacityProcess Control

Modularity

PrIME

Fraction Precipitation

Ion-ExchangeChromatographyAffinity Chromatography

Bayer selected PrIME as the technology of the future – requires:•5 years to achieve US FDA validation; and•US$100M to scale up to 3‐ 5 M litres capacity

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PrIME process modular scale up

10 L plasma per Separation Unit 250 L – 1000 L batch production

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• Delivery/Storage• Conditioning• Thaw• NAT Testing• Cryoprecipitate Removal• S/D Treatment• Equilibration

• PrIME• UF/DF• Anion/Viral Removal• Concentration, Diafiltration and formulation• Fill/Package• Store/Shipping

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Source Plasma

FiltrationCryo-supernatant Cryo-precipitate

SD viral inactivation

PrIMEIgG-depleted

plasma

Crude IgG

Polish

Nano filtration

UF+formulation+fill

PrIME

Crude Albumin

Polish

Nano filtration

UF+formulation+fill

IgG & Albumin-depleted plasma

AffinityChromatography

Factor IXconcentrate

SD viral inactivation

PrIME

Chromatography + Gel filtration

Nano filtration

UF+formulation+fill

Factor VIII or vWF

AlbuminIgG

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►The Singapore plant has previously been a cGMP certified by Singapore Health Science Authority. The plant was also previously audited by the FDA and EMA.

The incorporation of the PrIME plant will take 9 months to receive cGMP re‐certification.

►This significantly reduces the time required for the production of first CUP (15 months) and clinical trial (30 months) products.

Singapore Process Plant

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Singapore Processing Plant: Water for Injection plant

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Singapore Processing Plant: Quality Control Lab

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Singapore Processing Plant: Plant Corridor

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Singapore Processing Plant: 200L Bioreactor, Harvest and Permeate Tank

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The PrIME Technology addresses the acute shortage ofplasma drugs in Asia by processing the wasted

plasma and producing a safer, cheaper locally manufacturedproduct. PrIME achieves this by:

•Processing the Wasted PlasmaPlasma represents 45% of the cost of the current therapeutic

products; •High Product Yield, Small batch size & Short production runs

PrIME produces twice the end product in less time and smaller batches;

•Added Product SafetyPrIME removes viruses and Prions other systems can’t;

•Low Capital CostPrIME is a low capital cost process.

Summary

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• Dr Kailing Wang – Director of Science and Technology, PrIME Biologics• John Manusu – Managing Director, PrIME Biologics

Acknowledgements

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Documents

NuSep Presents at Bioplasma Conference2013/09/06 · Prakash Patel Managing Director +61 2 8415 7300 [email protected] About NuSep NuSep (ASX: NSP) is a publicly listed life