Dr Peter Levison of Pall Scale up of a Purification Platform for Manufacture of Biologics

Scale-Up of a Continuous Purification Platform for Manufacture of Biologics Peter Levison, Ph.D. Executive Director Business Development, Pall Biotech, Portsmouth, UK 3rd International Symposium on Continuous Manufacturing of Pharmaceuticals: Implementation, Technology and Regulatory, London 3rd October 2018 This presentation is the work product of Pall Corporation and no portion of this presentation may be copied, published, performed, or redistributed without the express written authority of a Pall corporate officer. Š 2018 Pall Corporation.

Pall Continuous Bioprocessing Solution from PD to Manufacturing PD to Manufacturing

Cell Culture Bioreactors

Clarification Cadence Acoustic Separator Cadence™ Acoustic Separator

Depth + 0.2 µm

Cadence Inline Concentrator (ILC)

Purification Capture – Cadence BioSMB + Cadence Virus Inactivation

2

Depth + 0.2 µm

Polish – Cadence BioSMB

Formulation Virus Filtration

Cadence ILC

Cadence Inline Diafiltration (ILDF)

0.2 µm

Large Scale Cadence Acoustic Separator Product Options

100 LPH Add-On 50 LPH Base Skid

3

350 LPH

Total Cell Density (x106 cells/mL)

Cadence Acoustic Separator – Scale Up Feed TCD

30 25

Stage 1

20 Stage 2

15 10

Stage 3

5

Stage 4

0 50 L/h

4.2 L/h Scale

50 L/h Scale

4.2 L/h Scale

Feed

Perm

% Red

Feed

Perm

% Red

TCD (106 cells/mL) Turbidity (ntu)

27.9 1496

2.1 251

92 83

31.6 1385

2.5 158

92 89

Viability (%)

81.5

n/r

81.5

n/r

Vol. Yield (%)

89

90 4

Scalable Continuous Chromatography

Process Development

Full Scale Cadence BioSMB Process 80 and 350

Cadence BioSMB PD Attribute

Patented Valve Cassette Design

Cadence BioSMB Process 80 Designed for perfusion applications with a single use flow path that can be set up in 30 minutes or less

Production time of 6 h for a 2000 L bioreactor with a single use flow path that can be set up in 30 minutes or less

3 – 16 *

3–8

3–8

0.6 – 3 L/h

10 – 80 L/h

30 – 350 L/h

Modular design Flexibility in Column arrangement (series or parallel) Number of Columns Flow Rates

Cadence BioSMB Process 350

Only multicolumn chromatography system that makes 24-hour process development a reality

* for ease of integration development 5

Cadence BioSMB Process 350 – Conditions Feed material: 400 L clarified cell supernatant with 5.8 gm/L mAb Chromatography sorbent: KANEKA KanCapA♦ PD Scale

Process Scale

4.0 mL/min

37 L/hr

11.2 mm ID x 5 cm H

14 cm ID x 5 cm H

Column volume

5 x 5 mL = 25 mL

5 x 0.77 L = 3.85 L

Residence time

3:04

3:04

5

5

46.4 gm/L

46.4 gm/L

Flow rate Column dimensions

Number of columns Operating binding capacity

> 150 x Scale-up 400 L processed in 13 cycles over 11 hours Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 6

♦ KANEKA KanCapA is a trademark of KANEKA Corporation

Process Consistency 13 cycles of operation Consistent UV elution performance Consistent impurity clearance

Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 7

Performance Comparison Batch Reference

BioSMB PD

BioSMB Process 350

Product conc.

9.54 gm/L

13.84 gm/L

13.85 gm/L

Product yield

98%

97%

97%

Aggregate

0.45%

0.72%

0.65%

LRV DNA

n.a.

4.2

5.0

LRV HCP

2.4

2.6

2.5

16 gm/L/hr

56 gm/L/hr

56 gm/L/hr

Spec.Prod.

Comparable process Cycles

3

13

Column diameter

40 cm

14 cm

Column height

16 cm

5 cm

20 L

3.85 L

Protein A volume

Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 8

Cadence BioSMB Process 350 System – User Data / Feedback A total of 2.3 Kg mAb (400 L at 5.8 g/L) was purified in 12 hours (13 cycles) Only 3.85 L of Protein A media was required (KANEKA KanCapA) A comparable batch process would require 20 L of Protein A media Five columns was the optimal configuration This is approximately 10% of the maximum capacity of the

Cadence BioSMB Process system Batch process was translated to continuous at PD scale and

transferred to Process Scale (150x) in less than 3 weeks Operation (HMI) of Cadence BioSMB Process system is

relatively easy (comparable with Cadence BioSMB PD) Result for Process Scale and PD Scale are very similar

(performance and product quality attributes) The Cadence BioSMB PD process successfully scaled up >150-fold 9

Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016

Cadence BioSMB Process 80 System – User Data / Feedback mAb process was translated from batch to continuous at PD scale and

transferred to Process Scale (40x) using a 4 column system

Productivity [g/(L*d)]

1000

800

600

400

200

0

Setting 1

Setting 2 Batch

BioSMB PD

Setting 3

BioSMB Process 80 Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018

10

Cadence BioSMB Process 80 System – User Data / Feedback A modified 3 column process was operated continuously for 10 days CQA’s remained consistent throughout the study Monomer content by SEC 100.0

Purity [%]

99.0 98.0 97.0 96.0 95.0 0

2

4

6

8

Day of experiment

11

10

Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018

Cadence BioSMB Process 80 System – User Data / Feedback A modified 3 column process was operated continuously for 10 days CQA’s remained consistent throughout the study 200

HCP [ppm] Res. Protein A [ppm]

Impurities [ppm]

160

120

80

40

0 0

2

4

6

8

Day of experiment

12

10

Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018

Cadence BioSMB Process 80 System – User Data / Feedback Consistent operation was reported across multiple cycles over 10 days UV Absorption [AU]

2.50 Cycle 01 Cycle 02 Cycle 03 Cycle 04

2.00 1.50

Cycle 05 Cycle 06 Cycle 07 Cycle 08 Cycle 09

1.00 0.50 0.00

Time in Cycle [hh:mm] 120.0

Conductivity [mS/cm]

Cycle 01

100.0

Cycle 02 Cycle 03

80.0

Cycle 04 Cycle 05

60.0

Cycle 06

40.0

Cycle 07 Cycle 08

20.0

Cycle 09

0.0

Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018

Time in Cycle [hh:mm]

13

Cadence BioSMB Process 80 System – User Data / Feedback Opportunities for cost reductions and process improvements

Save Time

Save Resin

Develop more projects

CTM

Lower ROI

More batches per year

Production

Higher throughput

Lower COGs

14

Faster Processes

Data presented by Ozan Ă–tes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018

Cadence Virus Inactivation System Comprised of one control unit and one or

two mixers Designed for both continuous and

batch operations Fully automated semi-continuous operation – For processing up to 2000 L fermentation volume – Single-use gamma irradiated flow path – GMP ready – Uses proven low pH virus inactivation methodology – Capacity up to 30 L/h volume

15

Cadence Virus Inactivation Operation Concept

16

Single-Use System Error free installation ensured by – Tagged connections – Shadow boarding – Unique genders and sizes on certain connections to make wrong assembly impossible

Mixer and manifolds can be installed in under

30 minutes Sample port in recirculation loop

for highly representative sampling No sample port on mixer

biocontainer

17

Cadence Virus Inactivation System

Phi 6 inactivation Start titer 1.33E+07 Cycle 1 2.00E+07 Cycle 2 NA Bag wash post cycle2 Bag control (no titration) 3.40E+07

End titer 0 0 0 3.30E+07

Integrated with Cadence BioSMB Process System, or standalone Single use flow path with installation in less than 30 minutes Capable of fully automated processing for batch chromatography No dead legs – Tested with: – Bacteriophage – B. diminuta – Riboflavin

Fully automated, designed for easy integration into continuous processes up to 2000L cell culture volume 18

Validation Results Test data available for testing with: – Bacteriophage (used as a virus analogue) to show effectiveness of the inactivation methodology – Bacterial challenge with Brevundimonas diminuta (B. dim) to show effectiveness of the design in eliminating hold-ups and carry over The bacteriophage data showed: – No active bacteriophages in the inactivation mixer and recirculation loop after completing two cycles, demonstrating the efficiency of the dosing, mixing and recirculation methodology – Controls showed no damage to the protein from pumping and mixing The B. dim testing data showed: – Complete washout of the target organism to below the limits of detection, indicating that the system does not have any detectable dead legs

19

Cadence BioSMB and Cadence Virus Inactivation Systems Together– Designed to be Integrated as a Combined Unit Operation Both systems can be integrated to run

continuously Automation – Connected directly or via control system to upstream and downstream operations

Process – Connected to Cadence BioSMB product port by interconnection manifold – Ability to collect whole elution peaks based on weight change in mixer or opening / closing of product valve

20

Evolution in Bioprocessing Key driver is time to market

Single-Use Technologies

Stainless Steel

Modular Construction

Continuous BioProcessing

First Movers

General acceptance

Second tier

Aging Technologies Adapted from: Morten Munk, PDA Annual Meeting 2016

Impact of Continuous BioProcessing Improved Facility efficiency Manufacturing flexibility

Reduced Processing time Operating costs Footprint Capital outlay 21

Detailed Commercial CoGs Breakdown: 20 batches / year at 6000 L and 5 g/L 16 Cost per Gram ($/g)

14 12 10 8

Other Labour Consumables Materials Capital

6 4 2 0 SS Batch

SU ICB SS Batch Platform Batch

Clarification

SU ICB SS Batch Platform Batch

Purification

SU ICB SS Batch Platform Batch

Polishing

SU ICB Batch Platform

Final Form.

ICB Platform exhibits 30% savings over SS Batch and 16% over SU Batch – $4.7M and $2.2M annual savings, respectively 22

Thank You peter_levison@pall.com