Impact of technology transitions on the plasma businessbo-conf.com/ppb09/present/papers/JO.pdf · commercialised by Sartorius Stedim Biotech: UVivatec ® Plasma Products Division

Plasma Products Division PPB09, Menorca, 11-14 May 2009

2-6-2009

Impact of technology transitions on theplasma business

Jan Over


2-6-2009

Technology transitions in thebiopharmaceutical business

Jan Over


Outline of presentation

• Trends in biopharmaceutical production

• New technologies- disposables- protein purification- virus and prion safety

• Recombinant / transgenic products

• Concluding remarks


Biopharmaceutical production:its present environment (1)

• pipeline of classic pharmaceuticals is drying up

• promise of protein therapeutics

• increase of protein products in pipeline


Biopharmaceutical production:its present environment (2)

• increase of regulatory requirements

• increasing cost

• cost-benefit analysis of therapeutics increasingly importantpressure on prices

• advent of biosimilars /generics

• increasing competition

• a financial turmoil going on


The regulatory environment

• robust processes

• Quality by Design

• DoE: Design of Experiments

• PAT: process analyticaltechnology

• Green by Design

• prion safety

http://courseweb.stthomas.edu/engineering/images/doe.gif


Trends in biopharmaceutical production (1)

• a clear need to be more efficient re time and cost,both in development and in routine manufacturing

• shorter development times shorter time to market

• higher expression levels in biotech (5 – 10 g/L)downstream processing will become a bottleneck ?

• higher throughput of 2nd and 3rd generation technologies

• alternatives to protein A for purification of monoclonals


Trends in biopharmaceutical production (2)

Cost reduction by:

- higher yields

- optimising capacity utilisation, minimise down-time

- flexible use of facilities, multi-purpose facilities

- more standardisation (but scalable), less validation

- lean manufacturing, less waste

- disposables


Disposables


A clear trend: disposables

Well established in the biopharmaceutical industry:

• tubings / connectors

• blood bags

• bags for buffers

• (large-scale) biocontainers

• clarifying filters, sterile filters (with or w/o filter housings)

• virus filters

• chromatography media (in batch mode)


Disposable chromatography ‘avant la lettre’

Sephadex turning 50 in 2009:

Since the early ’70s Sanquin is using DEAE-Sephadex A-50 as a disposable in its production of prothrombin complex concentrate….

http://images.google.nl/imgres?imgurl=http://i132.photobucket.com/albums/q27/redhot_553/happy-birthday.jpg&imgrefurl=http://www.myspace.com/liesbeth666&usg=__ApJhpntF85uRtVPW8EEZ6Xj0wag=&h=301&w=309&sz=19&hl=nl&start=19&tbnid=DVKaX68aEMEp1M:&tbnh=114&tbnw=117&prev=/images%3Fq%3Dhappy%2Bbirthday%26gbv%3D2%26ndsp%3D20%26hl%3Dnl%26sa%3DN


Disposables, general (1)

Advantages:

- no cleaning- no cleaning validation- no cross-contamination- increased flexibility- less capital investment, no maintenance cost- shorter processing times- shorter ordering / commissioning times- shorter time-to-market- lower classification of production area possible- less environmental effects ?



Lower water and energy usage: lower overall cost ?

Source: Rawlings & Pora,BioProcess International,March 2009



Some challenges:

- leachables and extractables- leakage / breakage of biobags- more suited to a certain scale (e.g. pilot and clinical trialproductions)

- incompatibility with some process fluids- incompatibility with extreme operating conditions



Points to consider:

- environmental impact (“green by design”)

- suitability for recycling

- waste disposal methods, waste management guidelines for specific components

- energy consumption

- cost-effectiveness


A disposable bioreactor system…

e.g. GE Healthcare Life Sciences,Sartorius Stedim Biotech

WAVE BioreactorTM


A biobag…

3 D Allegro,Pall Life Sciences

and a disposable system forbuffer and media preparation…

Mobius® FlexReady family


A disposable tangential flow filtration system…


A (disposable) chromatography system…

ÄKTATMready


Disposable processing, the near future?

http://images.google.nl/imgres?imgurl=http://www.healthpopuli.com/uploaded_images/GE_healthcare-794408.jpg&imgrefurl=http://www.healthpopuli.com/2007_12_01_archive.html&usg=__ghbRBnpbmDSRWuXAYoDqCYsg42s=&h=508&w=658&sz=82&hl=nl&start=1&tbnid=5L88AB7MjfdKhM:&tbnh=107&tbnw=138&prev=/images%3Fq%3Dge%2Bhealthcare%26gbv%3D2%26hl%3Dnl%26sa%3DG


Suppliers of disposable chromatography

- GE Healthcare Bio-Sciences

- Sartorius Stedim Biotech

- Upfront Chromatography

- ProMetic BioSciences

- Pall Life Sciences

- Tarpon Biosystems


Protein purification


Modern protein purification tools

• New ion-exchange media

• Membrane chromatography

• Multimodal chromatography matrices

• Expanded bed chromatography

• Affinity chromatography


New ion-exchange mediaCharacterised by:

- (very) high dynamic binding capacities (up to 200 mg/ml)- high flow rates (up to 600 cm/hour or higher, depending onbed height)

- similar characteristics as classical ion-exchangers

resulting in high productivity, high yields, reduced process time

Examples:- Capto series (GE Healthcare): Capto S, Capto Q, Capto DEAE,based on high-flow agarose modified with a dextran surfaceextender

- Q & S Hypercel (Pall), based on a cellulose matrix


Membrane chromatography- no bed packing- same dynamic binding capacity as conventional resins- short processing time (no diffusion limitation)- most suitable for large and unstable molecules- disposable- both for flow-through mode and bind-and-elute mode (inspecific cases)

- capable of virus removal (Q-membranes)

• Sartobind Phenyl membraneSartobind Q- and S-membranes (Sartorius Stedim Biotech)

• Mustang Q, S and E (Pall Life Sciences)


Multimodal chromatography (1)

Principle: ligands providing more than one mechanism of binding a protein.

Performance:- binding at higher ionic strength; for elution a change in pH may be needed beside a higher ionic strength.- higher selectivity compared to classical ion-exchangers

DoE is recommended to establish optimal processing conditions


Multimodal chromatography (2)

Examples:

- GE Healthcare, multimodal ligands for polishing purification of Mabs (Capto Adhere) or cation-exchange (Capto MMC)

- Upfront Chromatography, multimodal ligands for monoclonals and immunoglobulins

- Pall Life Sciences, multimodal ligand for monoclonals (MEP Hypercel) and other proteins (PPA HyperCel, HEA HyperCel)

- ProMetic, multimodal ligands for FVIII/von Willebrand factor, fibrinogen, immunoglobulin, albumin, ….


The ‘cascade’ concept of plasma fractionation

ProMetic’s Plasma Protein Purification System, using the Mimetic Ligand™purification platform.Licensed to Kedrion, Wuhan Inst. for Biol. Products and Blue Blood Biotech Corp.


Expanded bed chromatographyThe Rhobust® processing platform of Upfront Chromatography:

cross-linked agarose, derivatised with mixed-mode ligands, containing a high-density phase of tungsten carbide.

Capable of handling crude feedstock without clarifying prefiltration.


Affinity chromatography (1)Approach of BAC: affinity ligands based on derivatives of camelid antibodies: VHH (single domain) antibody fragments (CaptureSelect®); non animal-derived, stable to cleaning agents


Affinity chromatography (2)VIIISelect, for B-domainless recombinant factor VIII

IgSelect, for polyclonal human immunoglobulin

KappaSelect, for kappa light chains


Virus and prion safety


Viral risks, is the problem solved ?

SD, pasteurisation and small-pore virus filtration are here, but a few challenges remain….

• SD: removal from plasma

• pasteurisation: protein denaturation / yields

• virus filtration:- separation of parvovirus B19 and HAV from large molecular

weight proteins- filter capacities and protein yields- cost


SD removalA new matrix for SD removal:

SDR Hyper D®

- applicable for plasma and morepurified proteins

- high binding capacities forTriton X-100 and TNBP

- proteins are excluded from innerspace of beads

- high pressure possible, shortresidence times sufficient

- option of single use


Small-pore virus filters• Planova 15N and 20N (Asahi Kasei Medical)

New: BioEx module for IgG: LRV for PPV > 5.5 log10

• Ultipor DV20 (Pall): > 3 – 6 log10 removal for viruses ≥ 20 nm- constant flow and LRV independent of protein concentration (> 200

mg/ml)

• Viresolve Pro (Millipore)- LRV for parvovirus ≥ 4 log10, for MVM ≥ 5.2 log10,- high protein throughput (av. 6.9 kg/m2) in 4 hrs until75% flow decay

- no change in LRV at high flow decay (up to 90%)

• Virosart CPV (Sartorius Stedim) (20 nm): no change in LRV at high flow decay (up to 90%) (tested with PP7 in the presence of IGIV, BSA and Mabs); LRV > 5 log10 for PPV and MVM


New virus inactivation technology: UV-C

Initially developed and patented by CAF-DCF (Brussels);

engineered by Bayer Technology Services;

commercialised by Sartorius Stedim Biotech: UVivatec®


Efficacy of UV-C


Prion safetyvCJD transmittable by blood transfusion; also transmittable by

plasma products ?

Some (potential) measures:

- Specific filters for erythrocytes (P-CAPT™, MacoPharma/PRDT;Leukotrap, Fresenius/Pall; Asahi Kasei Medical)

- Positively charged depth filters for protein solutions (Pall)

- Specific filter for plasma and plasma products (PRDT, Allprion)

- Screening assay for prions (Amorfix, Allprion, Adlyfe, BioPeople)


Crucial aspects of a vCJD screening assay

- sensitivity (level of detection)

- specificity:assume a test specificity of 99,5% and a prevalence of infectious, but symptomless vCJD incubating donorsof 1: 10,000, then….testing 100,000 donors will yield 500 positive results of which10 will be true positives and 490 false positives….

In case no confirmatory test is available: what do we tell these donors ??


Sample# of

individual donors

# positive on initial screen

% false positives

# positive on retesting

U.K. fresh plasma 1,000 7 0.70 0/7

U.S. fresh plasma 322 2 0.62 0/2

France fresh+ frozen plasma 10,000 80 0.80 6/80

99.94%success on retest

99.8%UK specification

99.5%EU specification

EP-vCJD™ Performance: False PositivesSource: Amorfix Life Sciences


Status of the EP-vCJD screening assaySource: Amorfix Life Sciences

- sensitivity: 100% of a 106 diluted brain homogenate in blood

- specificity: at least 99.94%(6 positives in a French donor cohort of 10,000)

- more studies required using animal samples

- not validated yet on human vCJD samples

- no good confirmatory test available (PMCA* in development)

* protein misfolding cyclic amplification


Prion safety

Is prion removing filtration a better solution than testing?

- effective for all positive donations (also undetectable ones)

- false-positive donors do not need to be notified

- potentially cheaper


Prion removing filtration

PRDT’s prion affinity absorbent:

- effective for a range of infectious prion strains(incl. sCJD and vCJD)

- > 1.22 log10 reduction of endogenous infectivity (belowdetection limit)

- > 3 log10 reduction of exogenous infectivity- > 107 ID50 removal capacity per ml adsorbent- high flow rates possible- high protein yields- minimal/no impact on the therapeutic product- now being implemented by Octapharma for Octaplas®

http://www.redcross.org/


Recombinant / transgenic products


Recombinant alternatives to plasma products

On the market:

- factor VIII- factor IX- activated factor VII- activated protein C- antithrombin (transgenic)- albumin- thrombin


Recombinant alternatives to plasma products

In development:- (deletion mutant of) plasmin (Talecris)- von Willebrand factor (Baxter)- factor XIII- C1-esterase inhibitor (transgenic) (Pharming)- fibrinogen, tissue sealant- alpha-1-antitrypsin- polyclonal immunoglobulin (transgenic) (Hematech; Symphogen)- factor VIII with extended half-life (Baxter; Bayer; NovoNordisk)- factor IX with extended half-life (Baxter/Nektar; BioVitrum/Syntonix)- factor VIIa (a.o.) with extended half-life (CSL Behring/Novozymes)


And stem cell technology?

It may be feasible as a form of gene therapy…

and potentially as a route to alternatives for classical blood cell products….,

but it will not be a realistic approach for producing plasma proteins.


Concluding remarks


A ‘simple’ plasma fractionation process comprises….

- a capturing step (MM, EBA, monolith,…)

- a purifying step (affinity, MM, monolith, ….)

- a polishing step (membrane chrom., ….)

- two virus inactivation steps(SD, UV-C, 15 or 20 nm virus filtration, pasteurisation)

- a specific prion removal step (?)

- sterile filtration (+/- freeze-drying)

7 steps, each having 95% step yield ± 70% yield overall


Impact on the plasma fractionation business ?- Disposables:

Yes, increasing usage more flexibility, lower cost

- New chromatographic matrices:Yes, they will be adopted higher productivity, better selectivity

- Testing for prions:Maybe, only if a confirmatory test becomes available higher safetyPrion filtration:Probable higher safety

- Recombinant proteins:Yes, more will come more competition

- Stem cell technology:No, highly unlikely as replacement for plasma products no impact


Thanks to….- Amorfix Life Sciences (G. Adams)

- Asahi Kasei Medical (E. Kederer)

- BAC (I. van Gemeren, R. Clasen)

- GE Healthcare (L. Lundh, A. Harbers, K. Allmér, M. Gruvegard)

- Pall Life Sciences (O. Triebsch, M. Jansen)

- Progress Project Management and Engineering (8 former colleagues)

- Sanquin Pharmaceutical Services (P. van Mourik)

- Sartorius Stedim Biotech (W. Arts, S. Ray)

- Upfront Chromatography (R. Noel)


Last remarks

Apologies to all companies not mentioned by name

And:I do not have any commercial association to the companies mentioned but Sanquin….


Thank you for listening !

Documents

Impact of technology transitions on the plasma businessbo-conf.com/ppb09/present/papers/JO.pdf · commercialised by Sartorius Stedim Biotech: UVivatec ® Plasma Products Division