Presentation to

Engineering Conference InternationalIntegrated Continuous Biomanufacturing Conference

Implementing process closure and continuous processing into the modern biopharmaceutical facility

Oct 23, 2013 Castelldefels, Spain

THE BIOPHARMACEUTICAL INDUSTRY IS CHANGING

FINALLY!

THE SAFEST PROCESS IS CLOSED, WHERE THE ENVIRONMENT IS A PARAMETER WITHOUT CRITICAL ASPECT

WE HAVE THE TECHNOLOGY

ASME BPE, ISPE, BPOG Collaboration

STEP 1: DEFINING CLOSED

Closed Process Defined

• Environment does not represent a Critical Aspect (No Impact)

• Product is not exposed to the surrounding environment

• Additions and withdrawals performed in closed fashion

• Validated to show sufficient layers of protection against risk of contamination from the environment

• Risk of contamination is not mitigated by housing operation in bioburden-free or particulate-free environment

• Contamination of closed system = breach of integrity

NOTE: Closed Sterile (although sterile is one form)

ALSO: Closed Single Use Systems (although one form)

APPLYING CLOSED SYSTEM CONCEPTS:

THE INCREDIBLE SHRINKING FUTURE FACILITY

1980s Traditional Multiproduct Facility

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Seed Cell Culture

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Seed Cell Culture

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

e.g. 2 products

1980s Traditional Facility

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Seed Cell Culture

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Seed Cell Culture

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

2 X 3 production BRX Redundant centrifuges

Redundant downstream

Redundant upstream

Many silos Highly classified (to reduce “risk” of bioburden)

1980s Traditional Facility

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Seed Cell Culture

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Seed Cell Culture

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Product 1

Product 2

Product 1

Product 2UPSTREAM CELL (+) DOWNSTREAM CELL (-)

Shared Shared

Not new concept

Courtesy Scott Probst , Bayer, BPOG

TELL QA PEOPLE TO CLOSE THEIR EYES

AND PLUG THEIR EARS

REMINDER:

FOR THE REST OF US…

1980s Traditional Facility SharedMultiproductUnclassified!

And regulatory agencies know about this “secret”

EUROPE/ USA FDA (approx)

GRADE ISO CLASS IN OPERATION

MAX # PARTICLES

0.5 µ & LARGER / M3MAX # OF VIABLE MICROORGANISMS

(CFUs) / M3

A 5 3500 <1

N/A 6 35000 7

B 7 350000 10

C 8 3500000 100

D "9" 3500000 (at rest) 200

Risk Management

Eureka! Bioreactors are closed!

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Seed Cell Culture

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Seed Cell Culture

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

DECLASSIFY

REMOVE WALL

Solution for Successful Cell Culture:

AIRLOCKS NOT REQUIRED

Closed Media Containers

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Close the Bioreactor NOT cove corners!

Sterile filter clarified broth = briefly exposed

0.2µ filter after CENTRIFUGATION to close

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Corridor

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MALMedia Hold

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Logistical segregation

Sterile filter clarified broth = briefly exposed

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Corridor

Inoc

Inoc

PAL

PAL

PAL

PAL

MAL

MAL

ALALMedia Hold

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Airlocks no longer requiredDeclassified; HVAC segregation no longer required

When Environment = No Critical Aspect

Product 1 is closedProduct 2 is closed

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Corridor

Inoc

Inoc

PAL

PAL

PAL

PAL

MAL

MAL

ALALMedia Hold

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Physical segregation not required

If Environment = No Critical Aspect

Efficiency in Inoc Prep Likely:Cadence of 1 BRX every 2 days

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

CentrifugeCorridor

Inoc

Inoc

PAL

PAL

PAL

PAL

MAL

MAL

ALAL

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Corridor

Efficiency in Centrifugation Use LikelyBallroomMultipurpose Room

If Environment = No Critical Aspect

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Corridor

PAL

PAL

PAL

MAL

MAL

ALAL

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled UnclassifiedBSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Corridor

Single redundancy likely sufficient

If Environment = No Critical AspectSterile Filter all Buffers or Use < 24 hours

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Corridor

PAL

PAL

PAL

MAL

MAL

ALAL

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridorCorridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Use Protected Powder Additions and Closed Liquid Additions

If Environment = No Critical Aspect

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Corridor

PAL

PAL

PAL

MAL

MAL

ALAL

PAL PAL

PAL MAL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Close SystemsSanitize skidsBlind runs

If Environment = No Critical Aspect

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Corridor

PAL

MAL

MAL

AL

PAL MAL

PAL MAL

Corridor

Clean Equipment

Storage

CorridorCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC PAL

PAL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Use Single Use Systems

If Environment = No Critical Aspect

Buffer Hold

Purification 1 V+

Purification 2 V+

Men’s Locker Room

Women’s Locker Room Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Corridor

PAL

MAL

MAL

AL

Corridor

Clean Equipment

Storage

CorridorCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-Classified

UC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Media Hold

Purifica tion V-

PAL MAL

PAL MAL

Regulatory Concern

If closed

With Technology Improvements

Buffer Hold

Purification V+

Men’s Locker Room

Women’s Locker Room Inoc

Inoc

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Corridor

PAL

AL

AL

Clean Equipment

Storage

CorridorCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-ClassifiedUC

= ISO 8, Grade C

= Uncontrolled Unclassified

BSC

BSC

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

Purifica tion V-

PAL MAL

If concern overcome

If in line dilution usedIf cytocentric isolator used

If Environment = No Critical Aspect

Buffer Hold

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

Corridor

Corridor

AL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-ClassifiedUC

= ISO 8, Grade C

= Uncontrolled Unclassified

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K 15

K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

ISOL

Consolidated V+ / V-

Note fewer staff

Smaller volumes required

If Environment = No Critical Aspect

Buffer Hold

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

Corridor

Corridor

AL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

7

8

9

= ISO 7, Grade B5

= ISO 8, Grade C= ISO “9”, Grade D

= ISO 5, Grade A

e8

CNC= Controlled Non-ClassifiedUC

= ISO 8, Grade C

= Uncontrolled Unclassified

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K 15

K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

ISOL

If all SS: UC space above

If Environment = No Critical Aspect

Buffer Hold

Purification 1 V+

Purification 2 V+

Purifica tion V-

Men’s Locker Room

Women’s Locker Room

Inoc

Inoc

Seed Cell Culture

Equipment Wash

Cell Culture 1 Harvest and Clarification 1

Cell Culture 2 Harvest and Clarification 2

Weigh DispenseDispensed Kit StorageMedia Prep

Buffer Prep

Centrifuge

Corridor

Buffer Hold

Centrifuge

Seed Cell Culture

Corridor

PAL

PAL MAL

MAL

Inoc

Inoc

PAL

PAL

PAL

MAL

PAL

PAL

MAL

MAL

ALALPAL

MAL

PAL

MAL

PAL MAL

PAL MALPAL MAL

PAL PALPAL MAL

PAL MAL

PAL MAL

PAL MALPAL MALCorridor

Clean Equipment

Storage

CorridorCorridor

Clean Equipment

Storage

Corridor

Corridor

Corridor

BSC

BSC

BSC

BSC PAL

PAL

DFB DFB DFB DFB

MAL

MAL

PALMAL PAL MAL

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

CF-100 CF-100

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

15K

Media Hold

Media Hold

Downstream Equip Wash

Column Packing

Warehouse

Cell Bank

Dewar

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

CorridorAL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

ISOL

Buffer Hold

15K

15K

15K

15K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K9900 m2 5400 m2 + 380 m2 buffer hold UC

If Environment = No Critical Aspect

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

CorridorAL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

ISOL

Buffer Hold

15K

15K

15K

15K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K4000 m2 ISO 8, 1000 m2 ISO 7 500 m2 ISO 8

How might this look?

HOW CAN CONTINUOUS PROCESSING HELP SHRINK THE FACILITY?

Batch Continuous

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

CorridorAL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

ISOL

Buffer Hold

15K

15K

15K

15K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2KSmaller volumes & areas required

Batch Continuous

Mabs Model:

• Perfusion versus Batch Cell culture

• Continuous Harvest/Clarification (ATF, centrifuge, etc

• Continuous Capture (SMCC Protein A)

• SMB/SMCC where possible in multi-chrom operation

• Nanofiltration

• UF/DF & Bulk Fill

Batch Continuous

Requirements:

• All operations must be closed and synchronized for long term operations

• Cell Culture must be “bpsterile”

• Clarification cannot contaminate upstream (but…)

• Capture must be closed (but…)

• Any static hold steps must be “bacteriostatic”

• If continuous, UF/DF & Bulk Fill must be bacteriostatic

Batch ContinuousMetrics from benchmark projects and models:

• Assuming 5:1 lower titer in Perfusion versus Batch

• 15K batch BRX for 1000 kg protein

• 1K to 4K perfusion BRX for 1000 kg protein

• Harvest batch 20K in 4-6 hours after 12-14 days

• Harvest continuous 24/7 (1-3 BRX volumes/day)

5000 m2 batch 3000 to 3800 m2 continuous ( -24 to -40%)

Batch Continuous

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

CorridorAL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

ISOL

Buffer Hold

15K

15K

15K

15K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2KAssume 24% reduction

Assume no change

Batch Continuous

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Cell Culture

Weigh DispenseDispensed Kit Storage

Media Prep Buffer Prep

Centrifuge

CorridorAL

AL

Clean Equipment

StorageCorridor

Corridor

Corridor

ISOL

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

15K

15K

CF-100 CF-100

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

ISOL

Buffer Hold

15K

15K

15K

15K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

Buffer Hold

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Weigh DispenseDispensed Kit Storage

Media PrepBuffer Prep

CorridorAL

AL

Equipment Storage

Corridor

Corridor

Corridor

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

Further 1000 m2 space savings or more achievable

Closed + Continuous = Low Cost

Facility Cost

Less competitiveHigher costLess flexible

Lower costMore competitive

Highly flexible

Buffer Hold

Purification V+/V-Men’s Locker Room

Women’s Locker Room

Equipment Wash

Cell Culture Harvest and Clarification

Weigh DispenseDispensed Kit Storage

Media PrepBuffer Prep

CorridorAL

AL

Equipment Storage

Corridor

Corridor

Corridor

ISOL

DFB DFB DFB DFB

JC

JC

15K

15K

15K

15K

Column Wash &Packing

Warehouse

Cell Bank

Dewar

Media Hold

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

2K 2K 2K 2K 2K

Closing #1

Closed & Continuous

Project Risk

Thank You

CRB Contributors:Kim NelsonMark von StwolinskiPatrick Sullivan