Efficient Scale Up of Therapeutic Antibody Manufacturing ...
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Efficient Scale Up of Therapeutic Antibody Manufacturing Processes: Helping to Achieve Clinical and Commercial Milestones Abstract • Efficient and successful technology transfer to manufacturing plant is an outcome of a systemac approach to scale up. • The systemac approach depends crically on understanding of engineering parameters as well as scienfic and praccal knowledge. • Successful “first-me right” approach of scale up ensures mely achieving of clinical and commercial milestones and compressed me-to- market expectaons which eventually speed up the delivery of products to paents. • Challenges during scale up can be contributed due to technical, facility and equipment’s constraints. • At Syngene, we have been successfully demonstrang ‘first-me right’ technology transfer of mammalian (CHO up to 2000L, Hybridoma up to 500L) and microbial (up to 200L) processes to our GMP manufacturing plant. So far, we have achieved ten technology transfer without any failure or major technical glitches. • Our ‘first-me right’ technology transfer approach has resulted in significant reducon in cost by avoiding mulple engineering/technical batches and meeng important project mile stones. Technology Transfer Approach Assessing Confidence on Process: Decision Tree Systemac and Well Documented Implementaon Vivek Farkade, Lead Scientist, Biopharmaceutical MSAT, Syngene International Limited, Bangalore, 560099 INDIA. Email: [email protected] LinkedIn: https://www.linkedin.com/in/vivekfarkademsat/ Objective: • Successful tech transfer using ‘first time right’ approach Required information: • Process and analytical methods • Equipment, raw material & consumables • Product quality targets and specifications Methodology: • Risk based systematic approach by assessing incoming data and project requirement • Assessment provides transfer and scale up strategy based on the confidence on incoming data (both process & analytical) to meet specific objectives Collect & review inbound information and project objectives in context of internal capabilities & facilities Direct transfer at the intended scale of manufacturing, optionally with an intermediate scale engineering run Conduct experimental work to 1 Supplement information and/or 2 Tweak process or 3 Improve/Develop process Process reproduction or consistency batches at small and/or intermediate scale High Low Medium Confidence in meeting objectives with available information? Transfer to GMP plant at intended scale Does process meet ALL intended product specifications tested using the same analytical methods that are intended for transfer? Does experience with similar functionality equipment exists ? & Are equipment similar? HIGH confidence Of success: Process execution at scale With engineering batch(s) Has process been run by client at intended scale of manufacturing preferably 3 batches?* Is the process well described? Can process be implemented w/o major change? Is process robust and development history available? YES NO NO NO NO NO NO YES YES YES YES Q's are not strictly sequenal or binary. Judgement to be exercised. *In case of less than 3 batches data, the decision is based on other available data LOW confidence of success: • Process familiarizaon/reproducon development/opmizaon experiments >> process confirmaon (integraon batches) • Process robustness MEDIUM confidence of success: • Familiarizaon/adaptaon/reproducon at small scale and/or Intermediate scale batches Upstream and Downstream Scale Up Strategy Upstream: • Characterized bioreactors across scale • Maintained kLa and power per unit volume (P/V) for bioreactors at each scale • Maintained Eddy size (to minimize shear stress) • Engineering parameters are assessed and appropriate mixing and gassing strategy are defined Downstream: • Chromatography steps – with constant bed height. For non-linear scale up, residence me is maintained • Filtraon steps – volume or gram of protein per unit area and flow rate (LMH) are maintained • Mapping hold me data with equipment sizing • Connuous centrifuge opmizaon is performed during engineering batch Crical Upstream Scale up Parameters Geometric Comparison of Bioreactors @ Syngene • Bioreactor critical geometrical parameters (shaded in blue) are comparable at different scales • Selection of right bioreactor eases the scalability from lab to production Characterizaon of Bioreactors @ Syngene Incoming TTD (Process and Analycal) TTD review and gap analysis Evaluaon from scale up and transfer perspecve Approach document Execuon of transfer Performance review and summary Protocols / at scale TTD / batch records / analycal method documents as per idenfied approach Approach document describes gaps in technology & facility, compares process parameters and equipment between sending and receiving units, idenfies risk and provides appropriate migaons Scale independent parameters: Unchanged during scale up Scale dependent parameters (Engineering parameters) pH Temperature Dissolved Oxygen Culture Duraon Feeding Temperature and/or pH shiſt Inial VCD Power per unit volume: Impacts mixing and volumetric mass transfer coefficient Shear Stress: • Tip speed • Integrated shear factor (ISF) • Kolmogorov eddy size Volumetric mass transfer coefficient (kLa): Used as a measure of the aeraon capacity of a bioreactor and depends on • Agitaon rate • Impeller design • Aeraon rate To derive scale dependent parameters, bioreactors are completely characterized at Syngene Dimensions Unit 2L PD (Finesse/ Sartorius) 10L PD (Sartori- us) 50L PD (Thermo) (5:1 Dispos- able) 100L BMP1 (Thermo) (5:1 Disposable) 500L BMP1 (Thermo) (5:1 Dispos- able) 2KL BMP1 (Thermo) (2:1 Disposable) Total Volume (V T ) m 3 0.003 0.013 0.066 0.12 0.66 2.57 Working Volume (V L ) m 3 0.002 0.0102 0.05 0.10 0.50 2.00 Vessel Diameter (D r ) m 0.13 0.19 0.35 0.44 0.76 1.19 Vessel Height (H T ) m 0.225 0.475 0.8 0.953 1.52 2.30 Liquid Height (H W ) m 0.15 0.43 0.52 0.66 1.13 1.79 Overall H/D NA 1.73 2.50 1.9:1 1.9:1 1.9:1 1.9:1 Working H/D NA 1.15 2.24 1.5:1 1.5:1 1.5:1 1.5:1 Impeller type NA Pitched blade,45 Deg Pitched blade,45 Deg Pitch blade,45 Deg Pitch blade,45 Deg Pitch blade,45 Deg Pitch blade,45 Deg Number of impellers Nos 2 (3 blades) 2 (3 blades) 1 (3 blades) 1 (3 blades) 1 (3 blades) 1 (3 blades) Impeller Diameter (D i ) m 0.054 0.080 0.111 0.146 0.251 0.398 Ratio of Impeller to Vessel Diameter (D i /D r ) NA 0.41 0.41 0.32 0.33 0.33 0.33 Impeller power number NA 1.5+1.5 3.00 2.10 2.10 2.10 2.10 Sparger type NA Micro sparger 20 micron Micro sparger 20 micron DHS (0.178mm) and open pipe DHS (0.178 mm) and open pipe DHS (0.178 mm) and open pipe DHS (0.582mm) / Frit sparger 25 micron Agitator shaft angle Deg 0 0 19.6 16.5 16.5 19.6 Agitation range RPM 180-201 120 - 200 30-200 30-200 30-150 20-75 P/V range (Considering Final working volume) W/m 3 27-40 27-37 19-22 19-22 19-22 19-22 0 15 30 45 60 75 0 25 50 75 100 125 150 175 200 Kolmogrove eddy size,micron Agitation,RPM Eddy size 10L Eddy size 50L Eddy size 100L Eddy size 500L Eddy size 2KL 0 10 20 30 40 50 0 25 50 75 100 125 150 175 200 P/V,w/m3 Agitation,RPM P/V 10L P/V 50L P/V 100L P/V 500L P/V 2kL Power per unit volume 0 2 4 6 8 10 12 14 0.00 0.01 0.02 0.03 0.04 0.05 0.06 Volumetric mass transfer coefficient, h-1 VVM using DHS 2KL 500L 100L 50L Volumetric mass transfer coefficient Agitation is decided based on power per unit volume P=Np.ρ.N 3 .D 5 Flow rate are proposed to meet k L a requirement Optimized engineering parameters for different bioreactors: P/V value: between 20-30 k L a: ≥ 5.7 per h Shear stress (Eddy size): ≥ 15µ These engineering parameters at different scale are critical for successful scale up Case Studies: Convenonal verses Rapid Scale up Client provided a technical informaon package having: • Molecule details and its characteriscs • Upstream process • Downstream process Confidence analysis - Process Process Questionnaires Case Study 1 Case Study 2 USP DSP USP DSP Does process meet ALL intended product specifications tested using the same analytical methods that are intended for transfer? Yes No Yes Yes Is the process well described? Yes No Yes Yes Can process be implemented w/o major change? Yes No Yes Yes Is process robust and development history available? No No Yes Yes Does experience with similar functionality equipment exist? Are equipment similar? Yes Yes Yes Yes Has process been run by client at intended scale of manufacturing with n ≥ 3? No No No No Confidence Level Medium Low Medium Medium Case Study 1: Process Performance • Anbody Scale Up from 10L to 500L with Intermediate Scale Up • Comparable cell culture profiles with respect to VCC, viability and ter were obtained across different scales • Cumulave recovery is comparable at different process stages and scales (10L, 100L and 500L) and within the expected range • In-process and release tests met the respecve acceptance criteria/ expected ranges • Anbody Scale Up from 50L to 2000L w/o Intermediate Scale Up • Process was established at Syngene at 50L scale and further scaled up directly to 2000L scale without intermediate scale. • Comparable cell culture profiles with respect to VCC, viability and ter were obtained across different scales • Quality parameters met the respecve specificaon and data was observed comparable. Success Stories • Successful transfer from client laboratory to Syngene manufacturing • Process performance, in-process aributes and final product quality met expectaons • No failure of batches – Systemac approach and due diligence led to “First Time Right” transfer • Scienfic and praccal knowledge helps to manage technical challenges due to equipment differences, resulng in smooth scaleup • Syngene employs a decision-tree based confidence assessment of incoming technology for successful transfer and scale up Acknowledgment: Author expresses highest gratude and acknowledge the contribuon and support of Syngene and Client team who have contributed directly or indirectly during the course of this work. Case Study 1: Consistency batches for USP at 10L scale DSP development and consistency batches Scale up 10L → 50L → 100L → 500L Case Study 2: Process familiarization at 50L scale Scale up 50L → 2000L Based upon confidence, work plan was decided Molecule type Cell line Number of Tech transfer Inial scale Final scale Engineering batch success rate IgG1 CHO 4 10L Glass/50L SUB 500L to 2000L SUB 100% IgG1 Hybridoma 5 10L Glass 100L to 500L SUB 100% Anbody conjugate CHO 1 2L Glass 500L SUB 100% Clarification Capture Chromatography Low pH to Int. Depth Polishing Chromatography Nanofiltration UFDF 0.2 μ Filtration Process Step Process Recovery (%) Process Recovery (%) 0 20 40 60 80 100 120 10 L Scale 100 L Scale 500 L Scale 500 L Scale 50 L Scale 10 L Scale Age(Hrs) Titre 0 48 96 144 192 240 288 336 @Syngeneintl Syngene Interna�onal Limited Syngene Interna�onal Limited
Transcript of Efficient Scale Up of Therapeutic Antibody Manufacturing ...
Efficient Scale Up of Therapeutic Antibody Manufacturing Processes:Helping to Achieve Clinical and Commercial Milestones
Abstract• Efficient and successful technology transfer to manufacturing plant is an
outcome of a systematic approach to scale up.• The systematic approach depends critically on understanding of
engineering parameters as well as scientific and practical knowledge. • Successful “first-time right” approach of scale up ensures timely
achieving of clinical and commercial milestones and compressed time-to-market expectations which eventually speed up the delivery of products to patients.
• Challenges during scale up can be contributed due to technical, facility and equipment’s constraints.
• At Syngene, we have been successfully demonstrating ‘first-time right’ technology transfer of mammalian (CHO up to 2000L, Hybridoma up to 500L) and microbial (up to 200L) processes to our GMP manufacturing plant. So far, we have achieved ten technology transfer without any failure or major technical glitches.
• Our ‘first-time right’ technology transfer approach has resulted in significant reduction in cost by avoiding multiple engineering/technical batches and meeting important project mile stones.
Technology Transfer Approach
Assessing Confidence on Process: Decision Tree
Systematic and Well Documented Implementation
Vivek Farkade,Lead Scientist, Biopharmaceutical MSAT,Syngene International Limited, Bangalore, 560099 INDIA.
Email: [email protected]: https://www.linkedin.com/in/vivekfarkademsat/
Objective:• Successful tech transfer using ‘first time right’ approachRequired information:• Process and analytical methods• Equipment, raw material & consumables • Product quality targets and specificationsMethodology:• Risk based systematic approach by assessing incoming data and project requirement• Assessment provides transfer and scale up strategy based on the confidence on incoming data (both process & analytical) to meet specific objectives
Collect & review inbound information and project objectives in contextof internal capabilities & facilities
Direct transfer at the intended scale of manufacturing,
optionally with an intermediate scale engineering run
Conduct experimental work to
1 Supplement information and/or2 Tweak process or3 Improve/Develop process
Process reproduction or consistency batches at small
and/or intermediate scale
High Low
Med
ium
Confidence in meeting objectives
with available information?
Transfer to GMP plant at intended scale
Does process meet ALL intended product speci�cations tested using the same analytical methods that are intended for transfer?
Does experience with similar functionality equipment exists ? & Are equipment similar?
HIGH con�dence Of success: Process execution at scale With engineering batch(s)
Has process been run by client at intended scale of manufacturing preferably 3 batches?*
Is the process well described?
Can process be implemented w/o major change?
Is process robust and developmenthistory available?
YES
NO
NO
NO
NO
NO
NO
YES
YES
YES
YES
Q's are not strictly sequential or binary. Judgement to be exercised.
*In case of less than 3 batches data, the decision is based on other available data
LOW confidence of success:
• Process familiarization/reproduction development/optimization experiments >> process confirmation (integration batches) • Process robustness
MEDIUM confidence of success: • Familiarization/adaptation/reproduction at small scale and/or Intermediate scale batches
Upstream and Downstream Scale Up Strategy
Upstream:• Characterized bioreactors across scale • Maintained kLa and power per unit volume (P/V) for bioreactors at each
scale• Maintained Eddy size (to minimize shear stress)• Engineering parameters are assessed and appropriate mixing and gassing
strategy are defined
Downstream:• Chromatography steps – with constant bed height. For non-linear scale
up, residence time is maintained• Filtration steps – volume or gram of protein per unit area and flow rate
(LMH) are maintained • Mapping hold time data with equipment sizing• Continuous centrifuge optimization is performed during engineering
batch
Critical Upstream Scale up Parameters
Geometric Comparison of Bioreactors @ Syngene
• Bioreactor critical geometrical parameters (shaded in blue) are comparable at different scales • Selection of right bioreactor eases the scalability from lab to production
Characterization of Bioreactors @ Syngene
Incoming TTD (Process and Analytical)
TTD review and gap analysis
Evaluation from scale up andtransfer perspective
Approach document
Execution of transfer
Performance review and summary
Protocols / at scale TTD / batch records / analytical method
documents as per identified approach
Approach document describes gaps in technology & facility, compares process parameters
and equipment between sending and receiving units, identifies risk and provides
appropriate mitigations
Scale independent parameters: Unchanged during scale up
Scale dependent parameters (Engineering parameters)
pHTemperature
DissolvedOxygen
CultureDuration
Feeding Temperatureand/or pH shift
Initial VCD
Power per unit volume: Impacts mixing and volumetric mass transfer coefficient
Shear Stress:• Tip speed• Integrated shear factor (ISF)• Kolmogorov eddy size
Volumetric mass transfer coefficient (kLa): Used as a measure of the aeration capacity of a bioreactor and depends on • Agitation rate• Impeller design• Aeration rate
To derive scale dependent parameters, bioreactors arecompletely characterized at Syngene
Dimensions Unit2L PD
(Finesse/Sartorius)
10L PD (Sartori-
us)
50L PD (Thermo)
(5:1 Dispos-able)
100L BMP1 (Thermo)
(5:1 Disposable)
500L BMP1 (Thermo)
(5:1 Dispos-able)
2KL BMP1(Thermo)
(2:1 Disposable)
Total Volume (VT) m3 0.003 0.013 0.066 0.12 0.66 2.57
Working Volume (VL) m3 0.002 0.0102 0.05 0.10 0.50 2.00
Vessel Diameter (Dr) m 0.13 0.19 0.35 0.44 0.76 1.19
Vessel Height (HT) m 0.225 0.475 0.8 0.953 1.52 2.30
Liquid Height (HW
) m 0.15 0.43 0.52 0.66 1.13 1.79
Overall H/D NA 1.73 2.50 1.9:1 1.9:1 1.9:1 1.9:1
Working H/D NA 1.15 2.24 1.5:1 1.5:1 1.5:1 1.5:1
Impeller type NAPitched
blade,45 Deg
Pitched blade,45
Deg
Pitch blade,45 Deg
Pitch blade,45 Deg
Pitch blade,45 Deg
Pitch blade,45 Deg
Number of impellers Nos 2 (3 blades)2
(3 blades)1 (3 blades) 1 (3 blades) 1 (3 blades) 1 (3 blades)
Impeller Diameter (D
i)
m 0.054 0.080 0.111 0.146 0.251 0.398
Ratio of Impeller to Vessel Diameter(D
i/D
r)
NA 0.41 0.41 0.32 0.33 0.33 0.33
Impeller power number
NA 1.5+1.5 3.00 2.10 2.10 2.10 2.10
Sparger type NAMicro
sparger 20 micron
Micro sparger
20 micron
DHS (0.178mm) and
open pipe
DHS (0.178 mm) and
open pipe
DHS (0.178 mm) and open
pipe
DHS (0.582mm) / Frit sparger 25
micron
Agitator shaft angle Deg 0 0 19.6 16.5 16.5 19.6
Agitation range RPM 180-201 120 - 200 30-200 30-200 30-150 20-75
P/V range (Considering Final working volume)
W/m3 27-40 27-37 19-22 19-22 19-22 19-22
0
15
30
45
60
75
0 25 50 75 100 125 150 175 200
Kolm
ogro
ve e
ddy
size,
micr
on
Agitation,RPM
Eddy size 10L Eddy size 50L Eddy size 100L Eddy size 500L Eddy size 2KL
0
10
20
30
40
50
0 25 50 75 100 125 150 175 200
P/V,
w/m
3
Agitation,RPM
P/V 10L P/V 50L P/V 100L P/V 500L P/V 2kL
Power per unit volume
0
2
4
6
8
10
12
14
0.00 0.01 0.02 0.03 0.04 0.05 0.06
Volu
met
ric m
ass t
rans
fer c
oeffi
cient
, h-1
VVM using DHS2KL 500L 100L 50L
Volumetric mass transfer coefficient
Agitation is decided based on power per unit volume P=Np.ρ.N3.D5
Flow rate are proposed to meet kLa
requirement
Optimized engineering parameters for different bioreactors:
P/V value: between 20-30
kLa: ≥ 5.7 per h
Shear stress (Eddy size): ≥ 15µ
These engineering parameters at different scale are critical for successful scale up
Case Studies: Conventional verses Rapid Scale upClient provided a technical information package having:• Molecule details and its characteristics • Upstream process • Downstream process
Confidence analysis - Process
Process QuestionnairesCase Study 1 Case Study 2
USP DSP USP DSP
Does process meet ALL intended product specifications tested using the same analytical methods that are intended for transfer?
Yes No Yes Yes
Is the process well described? Yes No Yes Yes
Can process be implemented w/o major change? Yes No Yes Yes
Is process robust and development history available? No No Yes Yes
Does experience with similar functionality equipment exist? Are equipment similar?
Yes Yes Yes Yes
Has process been run by client at intended scale of manufacturing with n ≥ 3? No No No No
Confidence Level Medium Low Medium Medium
Case Study 1: Process Performance• Antibody Scale Up from 10L to 500L with Intermediate Scale Up
• Comparable cell culture profiles with respect to VCC, viability and titer were obtained across different scales
• Cumulative recovery is comparable at different process stages and scales (10L, 100L and 500L) and within the expected range
• In-process and release tests met the respective acceptance criteria/expected ranges
• Antibody Scale Up from 50L to 2000L w/o Intermediate Scale Up
• Process was established at Syngene at 50L scale and further scaled up directly to 2000L scale without intermediate scale.
• Comparable cell culture profiles with respect to VCC, viability and titer were obtained across different scales
• Quality parameters met the respective specification and data was observed comparable.
Success Stories
• Successful transfer from client laboratory to Syngene manufacturing• Process performance, in-process attributes and final product quality
met expectations• No failure of batches – Systematic approach and due diligence led to
“First Time Right” transfer• Scientific and practical knowledge helps to manage technical challenges
due to equipment differences, resulting in smooth scaleup• Syngene employs a decision-tree based confidence assessment of
incoming technology for successful transfer and scale up
Acknowledgment:Author expresses highest gratitude and acknowledge the contribution and support of Syngene and Client team who have contributed directly or indirectly during the course of this work.
Case Study 1:Consistency batches for USP at 10L scale
DSP development and consistency batches
Scale up 10L → 50L → 100L → 500L
Case Study 2:Process familiarization
at 50L scale Scale up 50L → 2000L
Based upon confidence, work plan
was decided
Molecule type Cell line Number of Tech transfer Initial scale Final scale Engineering batch
success rate
IgG1 CHO 4 10L Glass/50L SUB 500L to 2000L SUB 100%
IgG1 Hybridoma 5 10L Glass 100L to 500L SUB 100%
Antibody conjugate CHO 1 2L Glass 500L SUB 100%
Clarificatio
n
Capture
Chrom
atogra
phy
Low pH to
Int.
DepthPolis
hing
Chrom
atogra
phyNanofiltr
ation
UFDF
0.2 µ Filt
ratio
n
Process Step
Pro
cess
Rec
over
y (%
)
Process Recovery (%)
0
20
40
60
80
100
120
10 L Scale
100 L Scale
500 L Scale
500 L Scale50 L Scale10 L Scale
Age(Hrs)
Tit
re
0 48 96 144 192 240 288 336
@Syngeneintl Syngene Interna�onal Limited Syngene Interna�onal Limited
