Industrial Bioprocessing Technology

Module 10 – Pilot Facility and Process Automation

November 5, 2020

Instructor: Derek Choy

Topics

1. Process scale-up and challenges

2. Pilot scale manufacturing
3. Single-use manufacturing
4. Manufacturing automation
5. New technologies in manufacturing

2
Process Scale-Up

Purpose:

To make (a lot) more product that is identical to the original

intended formulation in terms of quality…

and do it in a robust and safe manner

3
Process Scale-Up
Success → Increase Demand → Increase in Production Scale
Motivation
• Increase total amount of material produced
(help more patients in need, more quickly)
• Improve productivity and efficiency
(reduce cost, wastes and environmental impact)
• Augment earnings
(enable/accelerate discovery and development of new drugs)
Challenges
• Reduced flexibility
• More attention to details and documentation required
• Higher risk (greater consequences from failure)
4
 Manufacturing Scale Bench Scale
Scale-Up – Cell Culture

5
 Scale-Up – Primary Recovery
Centrifugation Depth Filtration Microfiltration
Bench Scale
Manufacturing Scale

6
 Scale-Up – Chromatography
Chromatography Equipment Columns Buffers
Bench Scale
Manufacturing Scale

7
Process Scale-Up Challenges
• Dedicated procedures and equipment required to execute
operations that are trivial at small scale
• More time required for each unit operation
• Changes in unit operation performance
• Additional concerns over:
• Safety
• Facility fit
• Regulatory compliance
• Scheduling and material logistics

8
Process Scale-Up Challenges
50mM Tris pH7.5, 1L

+
6.057g Tris Base Several mL HCl
50mM Tris pH7.5, 1000L

+
6.057kg Tris Base Several L HCl
Need to use blended Tris Base and Tris-HCl Recipe
9
Safety at Manufacturing Scale
• Mass – Increases chances of injury from trying to move
heavy objects, or by objects moving into you
• Pressure – Any levels of pressure (even low pressure) can
become deadly when associated with high volumes
• Tripping/Slipping – More frequent and serious at scale
• Visibility – As processes and manufacturing plants grow
larger, each individual is less likely to be able to view the
entire operation, and is also less likely to be seen by others
• Spills/Leaks/Splashes – Releases of large volumes of even
water can have serious consequences
10
Facility Fit Considerations
• Availability of equipment at site
such as filter holders, chromatography columns, freezers…etc.
• Capability of equipment at site
such as maximum flow, pressure, number of cassettes…etc.
• Compatibility of equipment at site
with specific consumables, connections…etc.
• Physical limitations of site
such as size of entryways, amount of floor space…etc.
• Local laws and practices at site
such as holidays, site policies, indoor climate settings…etc.

11
Monte Carlo Simulations for Facility Fit Modeling
Repeated calculation of output variables with random input
variables constrained by a probability distribution
Load Volume(L) x Load Concentration(kg) x Yield(%) = Final Mass(kg)
Input Input Input Output

Randomized Input Variables (Titer, Step Yields…etc.)

Probability Distributions (Triangle, Normal, Discrete…etc.)
Output Variables (# of cycles or filters, final mass…etc.)
Reactor Titer (g/L) / Parameters Reactor Titer (g/L) / Parameters Reactor Titer (g/L) / Parameters
Comparison with Triang(6,7,8) Comparison with Normal(7,0.7) Comparison with Discrete({},{})
6.316 7.683 5.85 8.15 6.200 7.900

5.0% 90.0% 5.0% 5.0% 90.0% 5.0% 5.0% 90.0%

5.0% 90.0% 5.0% 5.0% 90.0% 5.0% 10.0% 90.0%
1.2 0.6 25%

1.0 Reactor Titer (g/L) / Parameters 0.5 Reactor Titer (g/L) / Parameters Reactor Titer (g/L) / Parameters
20%
Minimum 6.0149 Minimum 4.1856 Minimum 6.2000
Maximum 7.9854 Maximum 9.8901 Maximum 7.9000
0.8 0.4
Mean 7.0000 Mean 7.0000 Mean 7.0600
Std Dev 0.4083 Std Dev 0.7004
15% Std Dev 0.4271
Values 5000 Values 5000 Values 5000
0.6 0.3

Triang(6,7,8) Normal(7,0.7) 10% Discrete({},{})

0.4 0.2
Minimum 6.0000 Minimum −∞ Minimum 6.2000
Maximum 8.0000 Maximum +∞ Maximum 7.9000
Mean 7.0000 Mean 7.0000 5% Mean 7.0600
0.2 Std Dev 0.4082 0.1 Std Dev 0.4271
Std Dev 0.7000

0.0 0.0 0%
5

7
6

9
4
5.5

7.0

7.5

7.0

7.2

7.4

7.6

7.8
6.0

6.5

6.0

6.2

6.4

6.6

6.8
8.0

8.5

8.0
10

12
Worst-Case vs. Monte Carlo Facility Fit Modeling
Number of 4m2 VF filters required?
Facility currently only has holders for a maximum of 10 filter cassettes

• If we have HIGH titer, −∞

Number of 4m^2 Viral Filters / Outputs
10.50

HIGH yields AND HIGH 30%

98.4%

Protein A Column Bed Height: 25cm

1.6%

volumes for ALL steps: 25%

VF Loading (200L/m2)
Titer: 6-8g/L
>10 filters needed 20%
Num

15% Maximum

• What really is the Mean

Std Dev
Values

probability of that
10%

happening?
5%

0%
5
3

7
6

9
8
4

10

11

12

13
13
Achieving Facility Fit

Multiple ways to reduce number of VF filters required:

Number of 4m^2 Viral Filters / Outputs Number of 4m^2 Viral Filters / Outputs
−∞ 10.50 −∞ 10.50

100.0% 100.0%
30% 40%
Protein A Column Bed Height: 25cm Protein A Column Bed Height: 22cm
25% VF Loading: 225L/m2 35% VF Loading: 200L/m2
Titer: 6-8g/L 30%
Titer: 6-8g/L
20%
Numb Numb
25%
M

15% Maximum 20% Maximum

Mean Mean
Std Dev Std Dev
15%
Values Values
10%

10%

5%
5%

0% 0%
5

5
3

3
2

7
6

9
8

8
4

4
10

11

10

11
14
Process Scales
Bench Pilot Clinical Commercial

Bioreactor volume (L) <50 50 - 1000 1000 - 2000 >2000

Chromatography
<10 10 – 45 30 – 80 >80
Column Diameter (cm)

GMP Required? No Yes

Number of Projects 100s 10s 1s

Purpose Discovery and Material Material Drug product to

process generation for generation for treat patients
development animal trials and human trials
process
development

15
Pilot Scale
• Scale-up of in-development processes to confirm
performance and product quality
• Scale-down of commercial processes to troubleshoot issues
and improve performance
• Develop and test new technologies
• Generate material for development activities, including:
• Process development
• Toxicological and pharmacological studies
• Analytical assays development
• Drug formulation and material stability studies
• Process performance studies
16
What is Single-Use Manufacturing
• Uses consumables (such as bags and tubing) to hold,
transfer and process material, instead of reusable
equipment (such as tanks and piping)

Traditional Single-Use

17
Establishing and Maintaining Sterility
Traditional Stainless Steel Bioprocessing
• All equipment must be sanitized (usually with steam and/or
caustic) before and after use
• Most connections between equipment are made while potentially
exposing material to outside environment
Single-Use Bioprocesing
• All consumables are sterilized (usually via gamma irradiation) at
the time of their manufacture
• Consumables are designed to isolate their interior from the
external environment to maintain sterility
• Methods to connect elements aseptically via welding and sealing
18
Single-Use Manufacturing – Benefits at Pilot Scale
• Flexibility – Design of consumables are be more easily
changed to enable new capabilities
• Speed – No need for sterilization of tanks and piping
between campaigns and molecules
• Sterility – Ability to completely isolate the material from
the external environment during operations
• Cost – Significant reduction in initial capital cost and allows
for quick adaptation to new technologies
• Environmental Impact – Reduced total life cycle impact on
climate, health and resources
19
Single-Use Manufacturing – Benefits at Clinical Scale

Data from GE, 2018

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Single-Use Manufacturing – Ballroom Concept
Upstream Production Bioreactor

Harvest Filtration

Downstream

Seed Train Chromatography

Viral Inactivation
Viral Filtration
UF/DF
All unit operations can be in the same room
Formulation
(ballroom concept) if the entire process is
isolated from the environment

Image Courtesy of GE, 2018

21
Single-Use Technology – Bags
• Large bags (>50L) need to be
structurally supported with
totes, single-use mixers (SUMs)
or bioreactors
• SUMs and bioreactors provide:
• Mass reading via load cells
• Mixing (system depends on vendor)
• Fluid jacket for temperature control
• Connectivity for meters and probes
• Mass flow controllers (MFCs) for
bioreactor air spargers

22
Single-Use Technology – Bag Films
• Films are comprised of
multiple layers
• Nylon or polyester outer
layer used for durability
• Ethylene Vinyl Alcohol
(EVOH) used for gas
barrier
• Polyethylene commonly
used as inert contact
layer
23
Single-Use Technology – Bag Features
• Ports with tubing can be
attached to bags
• Clamps are often used
to block flow line
• Other features include:
• Impellers or other mixers
• Air sparger for bioreactors
• Probe ports (sometimes
with single-use probes)
• Sampling ports
• Pressure sensors

24
Single-Use Technology – Mixing Systems

Top Mount Impeller

Top Mount Wand Recirculating Flow
(eg. Thermo
(eg. Pall Wand Mixer) (eg. Saint Gobain)
HyPerforma)

25
Single-Use Technology – Mixing Systems

Bottom Mount Plunger Bottom Mount Impeller

(eg. Thermo ASI imPULSE) (eg. Pall Allegro)

26
Single-Use Technology – Mixing Systems

Bottom Magnetic Mount

(eg. GE Xcellerex,
Millipore Mobius,
Rocking Platforms Sartorius Palletank)
(eg. GE WAVE Bioreactor)
Air sparger integrated into bottom mount
mixer in GE Xcellerex XDR bioreactor
27
Single-Use Technology – Mixing Systems
Factors to consider when selecting mixing system:
• Amount of mixing achieved (time and power required to
reach homogeneity)
• Minimum working volume
• Shear generated
• Ease of installation
• Scalability
• Application specific considerations (such as fluid viscosity,
fluid type…etc.)

28
Single-Use Technology – Common Connections
Tri-Clamp (TC) Colder MPX/MPC Aseptic

Hose Barb

29
Single-Use Technology – Aseptic Operations
Tube Welder Tube Sealer
For connecting tubes aseptically For sealing tubes aseptically

Tubing material must be weldable/sealable: melt at high temperature

and reform into leakproof seal after cooling
30
Single-Use Technology – Moving Fluid
Peristaltic Pump Disposable Pump Heads

Pumpable tubing
required: pump
Quattroflow Pump
motion causes wear
on tube that may
result in structural
collapse over time

Levitronix PuraLev

Pump heads can be sterilized via

gamma irradiation
31
Single-Use Technology – Temperature Control

Chilled Water/Glycol Temperature Control Unit (TCU)

Temperature change for

large volumes will require
mixing for convective heat
transfer and will take
considerable time
Chilled TCU
32
Single-Use Technology – Disposable Flow Paths
Disposable Flow Path for Disposable Flow Path for
Chromatography Skid UF/DF Skid

33
Custom Consumables Design Guidelines
• Safety – The following factors should be considered for all
components on the custom consumable
• Operating temperature and pressure
• Welding and pumping requirements (tubing)
• Mechanical and physical properties required
• Compatibility – Consumables should have connections or
component sizes that can easily interface with other
consumables in the facility
• Flexibility – Additional functionality or space for expansion
to meet future needs without major redesign

34
Automation in Manufacturing
Lab Scale Manufacturing Scale

• One power plug • Multiple powered components

• One controller • Multiple controllers
• All parts within reach • Equipment spread across large area
• Standardized equipment • Unique custom components

35
Automation in Manufacturing
Programmable Logic Controller (PLC):
Controls single piece of equipment
(eg. Allen-Bradley)

Distributed Control System (DCS):

Controls unit operations with multiple
pieces of equipment
(eg. SIMATIC PCS 7, PlantScape or DeltaV)

Supervisory Control and Data Acquisition

(SCADA): Schedules and controls multiple
unit operations (possibly from different
plants) and output data management
36
Automation in Manufacturing

Operator Operator runs and interacts with recipe

Recipes Code for operating modules

Modules Logic for operating equipment

Equipment (may have its

Equipment own programming)

37
Automation in Manufacturing
• Continuous monitoring and control of all equipment in
plant (such as setting and responding to alarms)
• Ensures consistency in execution of process
• Allows the use of custom components and addition of new
capabilities in the future
• Historian records all output data, and assist in data review
and analysis for process troubleshooting or optimization
• Facilitates authoring and management of documentation
for compliance (such as electronic batch records)

38
Pilot Plant for New Technologies Development
New Automation Recipes
• Variable residence time loading
• UV controlled filtration
New Process Technologies
• Perfusion bioreactors
• Multi-column chromatography
• Integrated batch processing
Process Analytical Technologies (PATs)
• Continuous concentration monitoring
• Continuous product quality assays
39
Multi-Column Chromatography

ChromaCon Contichrom Cube GE ÄKTA pcc 75 Pall Cadence BioSMB PD

LEWA EcoPrime Twin GE ÄKTA process pcc Pall Cadence BioSMB Process 80/350
40
Multi-Column Continuous Capture – Scale-Up

Benefits:
• Reduced resin requirements
• Increased resin lifetime
utilization (clinical scale)
• Improved productivity
• Reduced buffer use
Challenges:
• New equipment required
• New recipes needed
• Extended processing time
• Scheduling
• Material stability

41
Integrated Batch Processing – Scale-Up

DS
V-2

V-2

Perfusion Harvest Capture VI Polishing VF UF/DF

(Multi-Column Continuous) (Pool-less Flowthrough)

Benefits: Challenges:
• Increased productivity • Start-up/shut-down
• Improved process consistency • Automation complexity
• Reduced facility requirements • New regulatory challenges
• Steady-state workload and • Process performance
materials flow • Batch definition
42
Online Process Analytical Technologies (PATs)
• Real-time measurements of process performance
• Allows collection of more data
• Supports on-the-fly changes in operation and long-term
process improvements
• Justify batch splitting for continuous processes
• Reduces staff workload and processing time
• Decreases chances of error and sampling contamination
• Enables new operational strategies

43
Online Concentration Measurement

Flow Fibrette
Flow Path
Teflon Seals

10mm Flow Cell

Images courtesy C Technologies, 2017

44
Online Analytical Chromatography

Depth
Filter V-2

UF / DF Drug
V-2

MCC Protein A Viral CEX / AEX Substance

Inactivation

HPLC/UPLC can measure:

• Titer (Analytical Protein A)
• %HMW (SEC)
• Charge Variance (ICF)

• Real-time titer information enable

immediate execution of capture step
• Real-time CQA measurements can:
• direct adjustments to operating conditions Waters PATROL
• justify batching in continuous processes
45
Summary
• Process scale-up involves safety, facility fit, regulatory and
logistics considerations
• Pilot plants play a critical role in scale-up and scale-down of
processes, as well as development of new technologies and
material generation for IND-enabling activities
• Single-use technology is useful at pilot and clinical scales
due to greater flexibility, and reduced risk and cost
• New technologies such as continuous processing and online
PATs can further improve productivity in manufacturing

46
Questions

47