Pragmatic implementation of single use technologies to deliver clinical supply

Pragmatic implementation of Single-use technologies to deliver clinical supply Priyabrata Pattnaik, PhD Director – Strategic Initiative

Agenda

Market and need assessment 1

2 PD Activities

Pilot Scale Run 3

4 Process Integration using Single use technology

Comparison of Bench and Pilot scales 5

6 Summary

Conclusions 7

MAb Market – Trends, Characteristics

mAbs ($48 bil) continue to represent the ‘growth’ driver for the biopharmaceutical market • Total ~ 1400 biologics projects in R&D and clinical phases • ~ 50% in preclinical and Phase I

Biomanufacturing capacity demand is ‘uncertain’ – Success rate of Ph1 to approval is <30%

– Not many easily accessible large manufacturing facilities

– A Flexible concept for approaching manufacturing is desirable

Significant capital investment required for commercializing a mAb – > 100-300 $ Mil installed cost for a traditional large scale SS production facility

Time to clinic is still a key driver especially for smaller and newer biotech firms

3

Data from Evaluate Pharma

4 4

“Commoditization” of mAb Processes

Developing a downstream (DSP) process for monoclonal antibody (MAb) purification is essentially a “solved” problem Time to clinic is still a key driver especially for smaller and newer

biotech firms Facilitated by template and single-use approach

“ … it is unlikely that non-conventional downstream unit

operations would be needed to replace conventional chromatographic and filtration separation steps, at least for

recombinant antibodies” - Brian Kelley, Biotechnol. Prog. 2007, 23, 995-1008

A pragmatic approach minimizes time and effort Template process Pre-select operating parameters to minimize PD work “Pre-package” devices/systems/ancillaries to reduce

specification, procurement and installation effort Use of ClinicReady Process Template Single-use technology

The Journey to Clinic Challenge - Reduce timelines with limited resources

5

6

Single-Use Technology in DSP of MAbs

Drivers Reduce/eliminate cleaning, utility and

validation costs Eliminate concerns of carryover Reduce turnaround time between

batches/campaigns Facilitate duplication of manufacturing

suites in multiple locations Ability to use same equipment with

various MAbs

Evidence for single use flow paths for entire DSP train has been sparse

Current Situation Single-use technology

most widely used in holding and preparing buffers Single-use flow paths for

certain unit operations at pilot scale

6

Technology & Product

Innovation

Knowledge & Expertise

What’s needed to produce material for clinical supply

Resources

Process Template Single Use Technologies

GMP Facility 7

8 8

CLARIFICATION AFFINITY

CHROMATOGRAPHY

ULTRAFILTRATION STERILE

FILTRATION

CEX CHROMATOGRAPHY

Bulk Drug

Substance

Bioreactor

VIRUS REMOVAL

ClinicReady MAb Process Template

AEX CHROMA-

TOGRAPHY

ProSep Ultra Plus Millistak+ Pod – D0HC / X0HC

Fractogel SO3

ChromaSorb

Viresolve Pro+ Express Pellicon 3

Ultracell

Process development space

Pre-select operating parameters

1 1 1 - 2 <

Protocols, Data collection, Analysis

tools & Scale up tools

+ X X # Unit

Operations X

# Unit Operations

Effort Risk

# Vendors # Devices options

# Process parameters

Execution Protocols? Data?

Analysis? Scale up? + X X

X

2 - 3 2 - 3 2 - 3 7

56 – 189 trials

14 trials 9

Proof of Principle

Bench Scale Millistak

D0HC + X0HC

P3 UltraCel

ChromaSorb

Fractogel SO3

ProSep Ultra Plus

e y

e y

Viresolve Pro+

Millistak X0HC

Template

10

[100L bioreactor]

Pilot Scale

Selection Tool

Sizing Tool

11

Two different mAbs – MAb04 and MAb08

11010090807060504030

30

25

20

15

10

HIC Elution Cond (mS/cm)

CEX

Elu

tion

Con

d (m

S/cm

)

MAb04

MAb08

UF/DF

Virus Filtration

AEX

CEX

Viral Inactivation

Protein A

Clarification

Unit Operation

Reduction of PD Parameter Space Fixed Operating

Parameter Flux

Residence Time

No pH adjustment to lower pH

Residence Time Constant pH

Flow Rate pH relative to pI

Conductivity range Flow rate

pH

Feed flux Diavolumes

Process time

Operating Parameter to be determined by PD

Capacity of depth filters Capacity of sterile filter

Capacity Elution buffer pH

Capacity of sterile filter

Capacity Elution Conductivity

Capacity

Capacity

TMP Concentration for DF

Operational Parameters to be established

Flux, Capacity

Residence Time, Capacity, Elution Buffer pH


Residence Time, Capacity, Loading pH, Elution pH and

conductivity

Flow rate, Capacity, loading pH and conductivity


Feed flux, # of diavolumes, concentration for DF, process

time

12

13

PD Parameter Space

UF/DF

Virus Filtration

AEX

CEX

Viral Inactivation

Protein A

Clarification

Unit Operation

Fixed Operating Parameter

100 LMH

3 minute Residence Time

No pH adjustment to lower pH

6 minute Residence Time Constant pH operation (5-5.5)

Flow Rate = 12.5 MV/min pH 1 unit below pI

Conductivity < 12 mS/cm Constant flux operation – 200

LMH pH 5 – 5.5

Feed flux = 5 LMM Diavolumes = 10

Process time = 3-6 hrs

Operating Parameter to be determined by PD

Capacity of depth filters Capacity of sterile filter

Capacity Elution buffer pH


Capacity Elution Conductivity

Capacity

Capacity

TMP Concentration for DF

Operational Parameters to be established

Flux, Capacity

Residence Time, Capacity, Elution Buffer pH


Residence Time, Capacity, Loading pH, Elution pH and

conductivity



Feed flux, TMP, # of diavolumes, concentration for

DF, process time

14

PD Data

Clarification – Secondary depth filter capacity

Virus Filtration

AEX

CEX

Protein A

Clarification – Primary depth filter capacity

Parameter

250 L/m2

~ 3 kg/m2

3 kg/L

68 g/L

58 g/L

85 L/m2

MAb04

374 L/m2

> 5 kg/m2

> 5 kg/L

> 100 g/L

45 g/L

300 L/m2

MAb08

150 – 400 L/m2

2 – 5 kg/m2

> 3 kg/L

> 50 g/L @ 6 min residence time

> 40 g/L @ 3 min residence time

50 – 125 L/m2

Expected Range

UF/DF 75 g/m2/hr

89 g/m2/hr 50-150 g/m2/hr

Note: In the case of MAb08, the cell culture process is a low titer, low density process. Hence, the depth filter capacities are higher than expected

Impurity Clearance – Bench Scale (MAb04)

Harvest Clarif iedHarvest

Protein A CEX AEX1

1000000

HC

P (p

pm)

LRV = 0.4

LRV = 2.8

LRV = 0.8

LRV = 0.9


Protein A CEX AEX


Protein A CEX AEXClarif iedHarvest

Protein A CEX AEX VF UFDF

Agg

raga

te %

Leac

hed

Pro

tein

A (p

pm)

0

4.5

1

1000000

DN

A (p

pb)

0

16

HCP < 10 ppm (ng HCP/mg MAb) DNA < 50 ppb (pg DNA/mg MAb)

< 10 ppm (ng HCP/mg MAb) < 2%

Disposable bioreactor for PD

16

Cell Growth similar between Small scale (3L, 50L) disposable or glass bioreactor (3L).

Viability is maintained and consistent whatever the scale

Productivity is comparable to stainless steel system

Scaling of Single Use Bioreactors

17

Mobius Single Use Chromatography

Mobius FlexReady Smart System: Modular & Automated

Smart Flexware Assembly:

18

Scale-up of developed Downstream Process Process from bench (~3-4 g) to 200L pilot scale (70-100g) using commercially available, off-the-shelf systems and single use assemblies

19

Depth Filtration Capture Step Virus Inactivation

Viral Filtration Tangential Final Filtration

AEX Chromatography CEX Chromatography

Last Step Drug Substance

First step Cell thawing

Cell amplification – 3 weeks Production – 2 weeks

7 weeks

USP week 1 to 5

DSP week 6

DSP week 7

Single Use - Process Scale (MAb04)

Bench Scale

100g process

0

10

20

30

40

50

60

70

Bench Scale 100 g

1.00E-011.00E+001.00E+011.00E+021.00E+031.00E+041.00E+051.00E+061.00E+071.00E+081.00E+09

Bench Scale 100g

DN

A (p

pb)

Leac

hed

Pro

tein

A (p

pm)

Protein A CEX AEX UF/DF

HC

P (p

pb)

1

1000000Bench Scale100g process

HC

P (p

pb)

1000000Bench Scale100g process

0

Agg

rega

te %

8

1000000

1

70

0

21

Charge Variants

min0 5 10 15 20 25 30 35 40

mAU

0

20

40

60

80

100

120

*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH CLARIFIED HARVEST1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH CEX POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH CHROMASORB POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH PROTEIN A POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\BENCH UFDF1.D)

UFDFProtein AChromasorbCEXClarified Harvest

min0 5 10 15 20 25 30 35 40

mAU

0

20

40

60

80

100

120

*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH CLARIFIED HARVEST1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH CEX POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH CHROMASORB POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\BENCH BATCH PROTEIN A POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\BENCH UFDF1.D)


Bench Scale Process Scale

min0 5 10 15 20 25 30 35 40

mAU

0

20

40

60

80

100

120

*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\PROCESS CLARIFIED HARVEST.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH CEX POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH CHROMASORB POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH PROTEIN A POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH UFDF1.D)


min0 5 10 15 20 25 30 35 40

mAU

0

20

40

60

80

100

120

*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\PROCESS CLARIFIED HARVEST.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH CEX POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH CHROMASORB POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH PROTEIN A POOL1.D)*MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH UFDF1.D)


min0 5 10 15 20 25 30 35 40 45

Norm.

0

5

10

15

20

25

30

35

40

MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\BENCH UFDF1.D) MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH UFDF1.D)

Bench ScaleProcess Scale

min0 5 10 15 20 25 30 35 40 45

Norm.

0

5

10

15

20

25

30

35

40

MWD1 A, Sig=280,16 Ref=360,100 (2011-09-13_WCX_TH\BENCH UFDF1.D) MWD1 A, Sig=280,16 Ref=360,100 (2011-09-01_WCX_MAB04\PROCESS BATCH UFDF1.D)

Bench ScaleProcess Scale

• Distribution of charge variants unaffected by unit operations at both scales

• Distribution of charge variants very similar in final pool from both scales

• Carboxypeptidase B digestion confirmed that basic peaks are same as C terminal Lysine variations

Comparison of Yields

Overall yield at bench scale and 100g scale ~ 85%

0102030405060708090

100

Bench Scale Process Scale

Yiel

d (%

)

22

Cost of Pilot Scale Runs - Summary Units Utilized

Hardware MIX sytems 2

Drum dollies 6

200L Bioreactor 1

Buffer systems 1

Chrom systems 1

Non-chrom systems [ CLF, VF, TFF ] 3 Systems/Hardware Cost ~ $2.0M

Disposables

MIX Bags 15

2D and 3D bags 22

Sterile filters 8

Devices 11 Single use flow paths 11

Total cost of disposables* ~ $50k 23 * Excludes chrom resins and TFF membranes

Resources at hand to produce the product for clinical supply

Upstream

Downstream

25

Summary

Demonstration of rapid scale-up of a MAb purification process using streamlined PD activities Eliminated screening multiple devices Minimized process development (PD) space by fixing certain

operational parameters Successful scale-up of entire downstream process using

commercially available, off-the-shelf, largely single-use systems and process containers Minimized engineering workload and start-up times by employing

pre-existing systems and assemblies

26

Thank you

Pragmatic implementation of single use technologies to deliver clinical supply

Technology

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