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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– 2 –
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
Restriction Membrane
Separation Membrane
Stream 2
Stream 1
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Stream 2
Stream 1
Restriction Membrane
Restriction Membrane
Separation Membrane
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Stream 2
Stream 1
Restriction Membrane
Restriction Membrane
Separation Membrane
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Sample Stream
Restriction Membrane
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%
Fact
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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
7 – Digested 10-2.1
8 – Digested 10-2.8
9 – Digested 10-3.5
10 – Digested 10-4.2
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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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01
2
34
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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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