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: Vivek.Farkade@syngeneintl.comLinkedIn: 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