inspiring advances in bioprocessing - repligen · •upstream continuous processing (perfusion)...

Inspiring Advances in Bioprocessing

John Bonham-Carter

March 14, 2017

Repligen Proprietary ǀ March 28, 2017 ǀ 2

Emerging trends, new challenges

• Upstream continuous processing (perfusion) proven to increase efficiency for 25+ years

• Integrating downstream continuous processing

Integrated continuous processing

• Time-savings driven by aggressive time-to-market goals

• Cost-savings

• Quality improvements

Shift to disposables

• Smaller footprint in smaller facilities

• Rapid growth in Asia

Regionalization

• COGS improvements – reduce capital expenditures

• Healthcare pressure costs

Focus on process economics

>40 on the market* and >350 in clinical development

Eight FDA approvals in 2014, 14 approvals anticipated 2015-2016

Robust pipeline to sustain future growth with superior efficacy, new targets, new disease areas

*Includes fusion proteins such as Enbrel and Eylea that use a mAb manufacturing process

Bioprocessing Trends


How Long Can Biologics Avoid Commoditization?

• Monoclonal antibodies may become commodities

– Biopharmaceuticals are not “unique” and cannot escape

the effects of the coming competition

– Penicillin price product cycle will probably not repeat – but

prices will fall over the next 20+ years

– Biosimilars, and multiple drugs for the same indication,

will impact cost strategies

• First-in-class biologics are not likely to be subject to

these cost pressures

– But not enough of these in development to support the

industry

• Future plans must include drugs with mid market sales

• Cost, flexibility and efficiency are the key focus moving

forwards

– New technologies being platformed in several new

commercial facilities

– Existing steel plants are being modified to boost

(“intensify”) production

Biotech? Pharma?

Biotech?


Upstream Production Options

“Perfusion culture manufacturing strategies for cell-culture derived biopharmaceuticals offer the potential of greater daily productivities and hence smaller facility footprints than batch and fed-batch culture manufacturing strategies..” Pollack 2013 Biotech and Bioengin 110: 206-219

Traditional

Perfusion

Concentrated

“For perfusion, you don’t really need exotic upstream equipment beyond robust cell-retention devices.” K. Konstantinov, Genzyme Corp, BioProcess International, December 2012

Leading cell separation device in perfusion MAb and rProtein production

Applications beyond perfusion - process intensification, cell banking, seed train optimization

In 8 of the Top 10 Pharma companies

In 7 commercial and 50+ Phase 1-3 processes

Award-winning filtration device

that delivers high cell

concentration and process

intensification

Technology

BPI Finalist of the Decade

2012


XCell Devices Utilize Alternating Tangential Flow (ATF)

0.2µM filter retains cells

inside bioreactor while

harvest is a clarified

stream ready for DSP

Connect the XCell System to a bioreactor. The system includes:

• A stainless steel housing comprising:

• single-use hollow fiber filter cartridge

• single use diaphragm pump

• A “ready to connect” single use device

Alternating Tangential Flow (ATF) is created within the device by the action of a diaphragm moving upward and then downward, powered by air within a pump head.

With each short 10 to 15 second cycle, a small back flush occurs at each end of the filter. This back flush prevents filter clogging and allows cell concentrations of over 300 million cells per ml.

OR

XCell Perfusion performance with high-density culture of Drosophila S2 cells


“Think continuous, think small”

What is it?

• A production method used

to manufacture and

process materials in a

consistent, constant, and

uninterrupted manner, in

contrast to batch

processing.

• Continuous upstream

processing (perfusion) has

been in use for over 25

years

Why is it important?

• Steady state operation

• Consistent product quality

• Reduced equipment size

• Increased productivity

• Streamlined process flow

• Low cycle times

• Reduced capital cost and

time

Needs and Trends

• Process intensification

• Scale-up – “Number up,

not scale up”

• Single-use technologies for

multi-product systems

• Impact on COGs

• Integrating upstream and

downstream


Continuous Biopharmaceutical Manufacturing: The Challenge…

7

Challenges

Science & Technology

Regulatory & Quality

Economic & Infrastructure

Organization & Leadership

Continuous Biopharmaceutical Manufacturing: The Challenge…Regulatory & Quality

Sharmista Chatterjee, CMC Lead for Qbd ONDQA / CDER/ FDA IFPAC Annual Meeting Baltimore, January 2012


Industry Experiencing Large Expansion

• ~30% XCell units go to seed train and N-1 perfusion

• ~30% people are specifying perfusion for clinical manufacture (Bioplan 2014)

• ~45% starting or expanding Continuous Processing (AspenBrook 2015)

Nothing New About cGMP Perfusion, Except Expansion

Fed-batch company converts after first trial “…The ATF gives us renewed interest in perfusion technology and how we can easily scale up to a commercial processes. The ATF is really enabling us to start imagining next generation processes…” Large East Coast Biopharma

Perfusion Expert “…The system is very convincing… it’s the Gold Standard for Perfusion…” Large European Biotech

Today, there are many thousands of trained

XCell perfusion engineers compared to hundreds a

decade ago

© 2016 Repligen Corporation. All rights reserved. The trademarks mentioned herein are the property of Repligen Corporation and/or its affiliate(s) or their respective owners.

Intensification Options

“Good”

What if you already have existing plants and little desire to change manufacturing platforms?


Large Volume, High Density, Cell Bag Directly Inoculates N-1 Perfusion Reactor

• Repligen’s high density XCell cell banking process was used to create 1L cryo bags

• A portion of these cells to inoculate a 1.5L wv bioreactor connected to an XCell™ ATF 2 perfusion system – mimicking a 200L (N-1) perfusion bioreactor

• An initial drop in the cell viability was expected as reported in the literature (Gargi et al. 2012, Wiley)

• Viability recovered as normal to produce a high viability high density seed culture for the N reactor

1 VVD

2 VVD

2.5 VVD


What Does N-1 Perfusion Achieve For Production Fed Batch?

• A 1.5L wv reactor was used to mimic a 2000L production reactor

• From the single N-1 XCell perfusion reactor, at 100E6 cells/mL, the cells were separately inoculated into low seed (0.5E6 cells/mL) and high seed (10E6 cells/mL) fed-batch production cultures

• The maximum VCD for high seed culture is 25E6 cells/mL and lasted only 7 days, whereas the maximum VCD for low seed culture is only 19E6 cells/mL and lasted 14 days

• The same protein quantity was produced in the high vs low seed cultures, but in about half the time


Shortening Vaccine Production Time with an Intensified Cell Bank

Ad35 TB Vaccine Candidate at Crucell (now Janssen Vaccines)

Source: 5th Bioman Pro Belgium 2008, M de Vocht, Crucell


Intensified Virus Production (iViP): Post Infection Perfusion

No change in VP/IU ratio compared to

standard process

Approx. 7.5x increase in volumetric

productivity


Crucell’s Supply Problem – 180MM doses required

Merck Achieved

Merck Projected

Bioreactor Size 1,000 L 10,000 L

Approx. doses per batch

0.25 MM 2.5 MM

Approx. doses per year

6 MM 60 MM

3 x 10,000 L reactors required

Traditional Process based on PER.C6 rAd 5 for Phase II HIV


Process Intensification And Perfusion Achieve a Remarkable Solution For Vaccine Manufacturing

Merck Achieved

Merck Projected

Crucell Achieved

Crucell Projected

Bioreactor Size 1,000 L 10,000 L 10 L 1,000 L

Approx. doses per batch

0.25 MM 2.5 MM 0.08 MM 8 MM

Approx. doses per year

6 MM 60 MM 1.98 MM 198 MM

3 x 10,000 L reactors required

1 x 1,000 L SUB sufficient

Intensified Process (and single use equipment)

Traditional Process based on PER.C6 rAd 5 for Phase II HIV



“Better”

How can you implement intensification and continuous processing to gain performance quickly?


Continuous Upstream Is Not Overly Complex – Once You Do the Work To Understand

Data from Veronique Chotteau

KTH Stockholm University

0

20

40

60

80

100

0

10

20

30

40

50

60

70

80

0 5 10 15 20 25

% V

iab

ilit

y (

Das

hed

Lin

es)

VC

D (

10

6 C

ells

/mL

)

Culture Time (days)

ATF2 Perfusion-MedDensity

Cell Bleed

0

5

10

15

0 5 10 15 20 25

Cu

mu

lati

ve Ig

G (

g)

Culture Time (days)

0

4

8

12

16

20

0 5 10 15

VC

D (

10

6 C

ells

/mL

)

Culture Time (days)

Fed-batch

0

5

10

15

0 5 10 15 20 25

Cu

mu

lati

ve Ig

G (

g)

Culture Time (days)


XCell™ ATF 6 - Stainless steel vs. Single-use formats

Identical performance, different configurations

XCell™ ATF 6 Stainless Steel

XCell™ ATF 6 Single-use

Parameters Stainless steel Single-use

Typical BR Volume (L) 50-200 50-200

ATF Pump Rate (LPM)

Nominal Max 20 20

Recommended Min 5 5

Recommended Max for Scale up 15-17 17

Filter Types and Surface Area (m2)

0.2um PS 2.1 2.1

0.2um PES 2.5 2.5

Pump/Filter Housing

Diaphragm Displacement Volume at 12 LPM (L)

1.5 1.5

Permeate Hold Up Volume (L) 1.3 1.8

Retentate Hold-up Volume (L) 2.2 2.2

Height (cm) 92.8 94.7

Weight of Dry Assembly (kg) 13.6 4.4


“Connect and Culture” – in as little as 30 minutes

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VCD: 80-100E6 Cells/mL BRV: 2.5VVD Flux: 5.7LMH Filter: PES 0.2um

• Similar growth profiles were observed between XCell™ ATF 2 SS and ATF 2 SU

Performance Comparison – XCell 2, SS vs SU

Beta Test Results


• Similar growth profiles were observed between XCell™ ATF 6 SS and ATF 6 SU • Connectivity, wetting and pre-use integrity procedures proven • Sterility maintained for 20 days of operation

VCD: ~ 40e6 cells/mL CSPR: 0.05nL/Cell/Day

BR w.v: 40L (50L SUB)

Beta Test Results

Performance Comparison – XCell 6, SS vs SU


• Similar cell specific productivity was obtained with XCell ATF 2 SS, ATF 6 SS and ATF 6 SU devices

• Demonstrates scale-ability of technology performance between ATF 2 and ATF 6 scales

• Demonstrates performance equivalence between ATF 6 stainless steel pump housings and ATF 6 Single-use devices

• Demonstrates compatibility of the XCell ATF Single-use device with Single Use Bioreactors

Beta Test Results

Cell Specific Productivity – XCell 2 & 6, SS & SU



“Better”

What if you have existing facilities – what advantage is there in adapting to an intensified process strategy?


Perfusion enables more process flexibility

Smaller, more flexible, more productive facility helps manage capacity

– In early clinical phases perfusion allows:

• Better utilization of reactor time

• Produce the same amount of product in a shorter time

• Product produced will have a more consistent quality

– In later clinical phases and scale-up

• Scale up in perfusion is lower risk because development scale is commercial scale

• In Fed-Batch the scale increases to 10-20x, which takes:

– more time

– additional steps

– additional process risk

– additional capital risk

Fed-Batch

Perfusion

200L

500L

5,000L

200L


Would You Retrofit Smaller Fed Batch Facilities With Perfusion Capability, Rather Than Expand To Larger Scale?

• Flexibility of manufacture: managing capacity with a 200L reactor

• In two weeks, FB produces 1kg;

• In a similar time, CP produces 0.45kg/day = 2.7kg (two 3 day batches plus wasted start-up time)

• If 4kg is required:

• Three further FB runs achieve goal on day 60 – or buy a 1000L reactor

• CP can be simply extended with same quality product, target reached day 16

Each batch provides 5g/L Each batch provides 1.5g/L at 1.5vvd

Which is riskier? Depends on your

existing knowledge…


Would You Build A New Large Scale $B Facility?

Would you run 75 SUBs in Fed Batch? Or 10x 15kL?

It will take 16-17 days for perfusion to make the same 50Kg and with the advantages of:

• 1000L SUB is used instead of 10kL Steel vessels

• Less time spent between reactor runs, less downtime

• Less cleaning and other non-productive efforts

• More constant daily operations, fewer variations

• Production can be more easily, and more quickly, expanded

• R&D scale is commercial scale, less scale up risk and late stage development time

Assume FB Titer at 5g/L 10kL provides 50Kg in one 14 day batch

Similar Qp, 2vvd perfusion may be 1.5g/L/day 1kL provides 9Kg per 3 day batch

Companies without the relevant skills need to improve their knowledge to

take advantage of these benefits

….Or maybe just 10 SUBs in Perfusion is preferable?

3-5 years to build?

1-2 years to build?



“Best?”

What if you could develop a new bioprocess platform based on state of the art techniques, leading towards real time release?


Where Can Process Intensification Make An Impact?

X X X

X X

Concentrating a process to produce

higher cell densities and higher product

yields in shorter time.

• Concentrated perfusion

• Concentrated fed batch

• N-1 or N-2

• Reducing the number of upstream

steps through increased cell

concentration at each step

• Cell Banking

• Cultivate large seed stocks and

eliminate seed steps


10L-50L 500L-2000L

Production:

Fed-Batch or

Continuous?

New, smaller, intensified manufacturing platform

• Significantly reduced capital required

• Smaller cleanrooms & facility, fewer reactors and utilities required

• Fewer operational steps, no open handling

• If run in continuous mode, automation provides for “same every day”

• Reduced tech transfer, changes dramatically the manufacturing risk and flexibility:

• Rapidity to build allows delayed facility build decisions

• Market uncertainty can be managed with copy/paste factories as needed

• Allows flexible geographic and CMO options, without risking capacity

Less scale up, saving time & cost

R&D scale is commercial scale

32 CONFIDENTIAL

• Small foot print

• Low residence time from protein expressed to protein in final formulation buffer – less degradation possible

• Low resin usage

• Just-enough production

• Low manual attendance need

• Continuous monitoring of the cultivation performance

• Integrated Continuous Bioprocessing Is Regularly Achieved At R&D Scale –

but not yet in cGMP production

End-to-end continuous production of complex recombinant proteins Tiainen P., Integrated Continuous Processing, 2013

Varied Drivers And Sophisticated Responses Are Driving Manufacturing Change…

Technology allowing more consistent, tighter, product quality

Process understanding

Availability of knowledgeable personnel

Demand for smaller, more productive facilities

Capital risks

Manufacturing and tech transfer risks

Manufacturing variability

Manufacturing CoG (becomes competitive advantage) Negative perceptions changing due to new reliable technology and increased knowledge

COGs is perhaps no longer the main driver for change. Each of these must be assessed for importance:

• Product Pipeline Mix

• Tech Transfer Risk

• Manufacturing Network Management

• Plant Flexibility

• Tighter Product Quality

• Existing Facility Utilization

Companies are responding to their specific needs with varied intensification solutions, and sometimes this is continuous processing

Intensification drivers

Continuous enablers

Managing the business and pipeline risk is now the key driver, not COGs, in the move to continuous bioprocessing


John Bonham-Carter

Director, Repligen

inspiring advances in bioprocessing - repligen · •upstream continuous processing (perfusion)...

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