Platzhalter Bild

“Downstream Processing 2.0:

From a Bottleneck to a Pacemaker”

ACS Biot

2010

Dr. Uwe Gottschalk, VP Purification Technologies, Sartorius Stedim

Biotech

What

are

the

hot Topics?

6th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan

Associates Inc.

6th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan

Associates Inc.

Finally

there

seems

to be

a Bottleneck

...

... depending

on who

you

ask

Who

is

facing

Limitations?

“Obviously there is no downstream bottleneck 

if you have unlimited cash”– K. John Morrow, Jr., PhD., 2009

Agenda

1.Improving

throughput

–

The

Capacity

Disconnect

1.Improving

throughput

–

The

Capacity

Disconnect

2.Process

Economy –

Cost

of Goods

matter

2.Process

Economy –

Cost

of Goods

matter

5.The Future of DSP –

Revisiting the Past

5.The Future of DSP –

Revisiting the Past

4.Benchmarks of the Future –

No best, just better

4.Benchmarks of the Future –

No best, just better

3.Discovering new Shores -

Fortune favors the Brave

3.Discovering new Shores -

Fortune favors the Brave

Data adapted from: F. Wurm

Production of recombinant Protein Therapeuticsin Cultivated Mammalian Cells. Nature Biotechnology 22, 1-6 (2004)

Improving Titres Improving Titres –– MAbsMAbs (Bulk API)(Bulk API)

Titre ImprovementsImportant cost benefits as titres go beyond 1g/LThese diminish as we go beyond 5g/L

Beyond 5g/LDownstream cost dominatesIn this example the plateau is just under $100/g

Challenge in DSPBioreactors decrease in size?Cope with increased titresNeed to drive out costs Implications for facility design Results from Biopharm Services Generic MAb cost model

Bulk API Direct manufacturing costs

0

100

200

300

400

500

600

700

800

0 2 4 6 8 10 12

Titre (g/L)

CO

G ($

/g)

2000L 5000L

4 Bioreactors

Estimate of CoG based on standard MAb process for bulk drug substance

A universal cost model for single use systems in biomanufacturing. Andrew Sinclair, BioPharm

Services; Berlin Oct. 2007

Hot Topic: High Titer

Processes

Jim Davis, Lonza

Economics of Monoclonal Antibody Production: The relationship between upstream titer and downstream costs; IBC San Diego March 2008

DSP is

Mass

not

Volume

driven

Platzhalter Bild

1.Improving

throughput

–

The

Capacity

Disconnect

1.Improving

throughput

–

The

Capacity

Disconnect

2.Process

Economy –

Cost

of

Goods

matter

2.Process

Economy –

Cost

of

Goods

matter

Agenda

5.The Future of DSP –

Revisiting the Past

5.The Future of DSP –

Revisiting the Past

4.Benchmarks of the Future –

No best, just better

4.Benchmarks of the Future –

No best, just better

3.Discovering new Shores -

Fortune favors the Brave

3.Discovering new Shores -

Fortune favors the Brave

Technical challenge

Sensitive and technically demanding products require processes with inherent

complexity and expensive infrastructure

Need for robust & scalable processes for the entire DSP

Increasing regulatory scrutiny (Comparability!)

Financial challenge

Processes are fixed-cost driven (Investment vs

Consumables)

Manufacturing costs 15 -

25% of sales price

Costs for DSP up to 75% of manufacturing costs

Cost Balance Benefit for innovative treatments

Biosimilars

Challenges

of a Modern Downstream

Process

Bringing

Biosimilars

to the

Market

Biosimilars: Margin

Squeeze

translate

into

lower

COGS

MAb

manufacturing: 6 x 2,000L tanks, 2g/L, 90% utilisation; 211 #/a; 527 kg/yr; invest 172 Mio Euro;

142 $/g; Sinclair 2006

Category

Typical

COG breakdown

by

Hot Topic: Use

of Disposables

J. Zhou

BPI Vienna 2008

Agenda

2.Process

Economy –

Cost

of Goods

matter

2.Process

Economy –

Cost

of Goods

matter

5.The Future of DSP –

Revisiting the Past

5.The Future of DSP –

Revisiting the Past

4.Benchmarks of the Future –

No best, just better

4.Benchmarks of the Future –

No best, just better

3.Discovering new Shores -

Fortune favors the Brave

3.Discovering new Shores -

Fortune favors the Brave

1.Improving

throughput

–

The

Capacity

Disconnect

1.Improving

throughput

–

The

Capacity

Disconnect

Downstream

Processing

1980

“If it ain’t

broke, don’t fix it”– Bert Lance, 1977

Downstream

Processing

2010

“Le mieux

est

l’ennemi

du bien”(better is the enemy of good)

– Voltaire, 1772

“没有最好，只有更好”(No best – only better)

– Chinese Movie Cliche

Companies

are

questioning

current

Standards

6th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan

Associates Inc.

Gelfiltration

CHO

Human

Hybridoma

BHK

Cell Removal/Clarification

Cell Removal/Clarification CapturingCapturing Intermediate

Purification

IntermediatePurification PolishingPolishing Virus

Clearance

Virus ClearanceFermentationFermentation

Microorg.

New process trainNew process train

X-FLow

Depthfilter

Centrifuge

CEX

Mixed-Mode

Protein A

HIC

Ceramic HA

CEX

AEX (B/E)

AEX (FT)

AEX-M

Size-Exclusion

Adsorption

Inactivation

EBA

K. KonstantinovBayer Corp, USA

CHO

Protein A

CEX

AEX-Membrane

New process train ready

CHO OrthogonalCentr./DF Protein A CEX AEX (FT)

Depthfilter

Centrifuge Inactivation

Adsorption

Size-Exclusion

Off the

Shelf

Platform

for

Rapid Process

Train

Assembly

Page 26

Unprocessed Bulk

Viral Filtration

CEX

Chromatography

AEX Chromatography

Mixed Modechromatography

Viral Inactivation

3 columns

Unprocessed Bulk

Viral Filtration

CEX

Chromatography

AEX Membrane chromatography

Mixed Modechromatography

Viral Inactivation

Unprocessed Bulk

Viral Filtration

CEX

Chromatography

HCICChromatography

Viral Inactivation

2 columns +

1 membrane 2 columns

1 column +

1 membrane

Unprocessed Bulk

Viral Filtration

CEX

Chromatography

AEX Membrane Chromatography

Viral Inactivation

Alahari

2009

Medarex: Non-Protein A based Purification Processes: Scheme Evolution

A Consensus –

Value

Chain in Bioseparation

Increasing biomass and contaminant levels

Protein A pool volumes and step cost

DNA & HCP levels post Capturing

Polishing load volumes and conductivity

Pathogen clearance as a moving target

High Titer

Implications:

Agenda

1.Improving

throughput

–

The

Capacity

Disconnect

1.Improving

throughput

–

The

Capacity

Disconnect

2.Process

Economy –

Cost

of Goods

matter

2.Process

Economy –

Cost

of Goods

matter

5.The Future of DSP –

Revisiting the Past

5.The Future of DSP –

Revisiting the Past

4.Benchmarks of the Future –

No best, just better

4.Benchmarks of the Future –

No best, just better

3.Discovering new Shores -

Fortune favors the Brave

3.Discovering new Shores -

Fortune favors the Brave

•

New generation

of lenticular filtration

media

•

No Diatomeaceous

Earth; Synthetic

•

Cell

removal, clarification

& early

on contaminant

removal

Biomass

Removal and Early

Contaminant

Clearance

Increasing biomass and contaminant levels

DNA & HCP levels post Capturing

addresses:

The

Focus on Column

Chromatography

is

increasing

6th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan

Associates Inc.

BioPharm

Intl. October

2007

John Curling and Uwe Gottschalk

Packed Bed Chromatography: The Good The Bad and The Ugly

U. Gottschalk. Bioseparation

in antibody manufacturing: The good, the bad and the ugly.Biotechnol

Prog. 2008 May-Jun;24(3):496-503.

Pete Gagnon

2007

Chromatography Technologies for DSP

Polishing

(Membranes)

• Highly porous structure

• Pore size: 3 –

5μm

• Convective Flow

• Minimal buffer useCapturing/IP

(Resins)

• Bead size distribution: 15 -160 μm

• Average pore size: 15 -

40 nm

• Diffusion limited flow

• High capacity

Hot Topic: Cost

of Capturing

6th Annual Survey of Biopharmaceutical Manufacturing. Eric S. Langer, BioPlan

Associates Inc.

D. Low BioManufacturing

Paris 2007

Protein A pool volumes and step cost

addresses:

Two Birds –one Stone: Contaminant Precipitation at Pfizer

Protein A pool volumes and step cost

DNA & HCP levels post Capturing

addresses:

Three Birds –

one Stone: Contaminant Precipitation at Medarex

Precipitation of Process-Derived Impurities in Non-Protein APurification Schemes for MAb; J. Wang et al. BioPharm

Intl. 10/2009, 2-9

CEX

TFF

Q Membrane2 g/ml

Dilution

HCP < 1000 ng/mg

VF

Mix Mode

Fig 7b. TFF based process

Dilution

HCP < 1000 ng/mg

CEX HCP < 10ng/mg

Contaminant precipitation

Q Membrane20 g/ml

Fig 7a. precipitation based process

VF

HCP BDL

Dilution

CEX

TFF

Q Membrane2 g/ml

Dilution

HCP < 1000 ng/mg

VF

Mix Mode

Fig 7b. TFF based process

Dilution

HCP < 1000 ng/mg

CEX

TFF

Q Membrane2 g/ml

Dilution

HCP < 1000 ng/mg

VF

Mix Mode

Fig 7b. TFF based process

Dilution

HCP < 1000 ng/mg

CEX

TFF

Q Membrane2 g/ml

Dilution

HCP < 1000 ng/mg

VFVF

Mix Mode

Fig 7b. TFF based process

Dilution

HCP < 1000 ng/mg

CEX HCP < 10ng/mg

Contaminant precipitation

Q Membrane20 g/ml

Fig 7a. precipitation based process

VF

HCP BDL

Dilution

CEX HCP < 10ng/mg

Contaminant precipitation

Q Membrane20 g/ml

Fig 7a. precipitation based process

VFVF

HCP BDL

Dilution

Protein A pool volumes and step cost

DNA & HCP levels post Capturing

Polishing load volumes and conductivity

addresses:

Hot Topic: Polishing load volume/conductivity

•

Salt Tolerant Interaction Chromatography

(STIC)

•

New cellulose-based membrane

structure

•

Primary

instead

of quartenary

amine

ligand

Polishing load volumes and conductivity

addresses:

Results

so far achieved:

No CHOP-breakthrough

at 10 kg MAb/ L of membrane

> 4.96 LRV for

MMV at 150m mM

NaCl.

Downstream Costs: Why bother?

Jim Davis, Lonza

Economics of Monoclonal Antibody Production: The relationship between upstream titer and downstream costs; IBC San Diego March 2008

Limitation: Oleosin yields < 1kg/ha

2000: Oleosin

Platform

Will Protein A Capturing ever be Single-Use?

2005: TMV Nanoparticles

Immunoabsorbent

nanoparticles

based on a tobacco mosaic virus displaying protein AS. Werner et al. PNAS 103, 17678 -

17683

Polyester Granule100-300 nm

Grage, K. and Rehm, B.H.A. (2008) Bioconj. Chemistry, 19(1):254-62.

Polyester Synthase

2010: Bio Polyester Platform

Hot Topic: Pathogen Safety

15. October 2008 Seite 2

UVC Inactivation

Nanofiltration (20nm)

Membrane Chromatography

Orthogonal Contaminant Removal Technologies

Depth filtration

Pathogen clearance as a moving target

More

Mileage

out of Virus Filtration

The problem: Virus filters shows low Vmax

and Flow Rate for blocking Feed Streams

Vmax2

Vmax

200 –

1000 L/m²50 -100 L/m2.h.bar

low or moderatly

blockinge.g. MAb

Vmax

<100 L/m²

5 -

30 L/m2.h.barhighly blockinge.g. IVIG, Enbrel

Agenda

1.Improving

throughput

–

The

Capacity

Disconnect

1.Improving

throughput

–

The

Capacity

Disconnect

2.Process

Economy –

Cost

of Goods

matter

2.Process

Economy –

Cost

of Goods

matter

5.The Future of DSP –

Revisiting the Past

5.The Future of DSP –

Revisiting the Past

4.Benchmarks of the Future –

No best, just better

4.Benchmarks of the Future –

No best, just better

3.Discovering new Shores -

Fortune favors the Brave

3.Discovering new Shores -

Fortune favors the Brave

The

Renaissance of Protein Purification

Michelangelo de Lodovico

Buonarotti

•

Centrifugation

•

Extraction

•

Precipitation

•

Filtration

•

Crystallization

•

UV-Inactivation

Old Enabling Technology: Boring but Reliable

“The

real voyage

of discovery

consists

notin seeking

new

landscapes

but

in having

new

eyes.”

Marcel Proust. 

Downstream

Processing

2010+

Yes

we

can!

Uwe.Gottschalk@sartorius- stedim.com

Thank

you!