elevator pitch: engineering car t cells on the...


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October 5, 2016 CliniMACS Prodigy CAR-T Cells


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Conventional genetic modification of T cells requires...

Donor /

patient

+ + ++ ++

Blood

LeukapheresisStorage

Application

Cell

selection

Cell

transduction

Cell

formulation

Cell

activation

Cell

expansion

CliniMACS Prodigy CAR-T Cells October 5, 2016


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Scalable

Automated

Standardised

Easy Tech Transfer

Cost Efficient

What Miltenyi Provides

October 5, 2016

GMP Raw Materials

LentiviralVectors

QC Instruments

and reagents

Prodigy

Cell processor

Training

CliniMACS Prodigy CAR-T Cells

The complete

CAR- T

package


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Magnet Unit

Liquid Sensor

Pinch valves

Peristaltic pump

Bag hangers

Bag

Compartment

CentriCult Unit

Touch screen

Gas Mix Unit Tube Sealer

Bar code reader

CliniMACS Prodigy

October 5, 2016October 5, 2016 CliniMACS Prodigy CAR-T Cells


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Prodigy: Fully automated clinical transduction

1. Guided installation of TS520 sterile tubing set

2. Sterile docking to CliniMACS® Prodigy:

• Reagents

• Patient sample

• Sterile connection for vector

3. Pre-transduction Cell processing

• Cell washing

• CliniMACS cell separation

4. Addition of TransAct activation reagent

5. Transduction with Lentiviral Vector

6. Cell cultivation

• Media addition and removal

• Supplement addition

7. Final product formulation

• Change buffer

• Concentration

8. Harvest ready to give to patient/cryopreserve



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TCT Process Software – In Built Flexibility

Guided User interface (GUI) for culture: “Activity matrix”

• Possibility to start a run at various steps

(e.g. Include/skip enrichment, culture

only, resume culture)“

• Flexibility of the manufacturing run

attained through user defined “activities”

• Transduction

• Shaking conditions

• Feeding (Feed or Media exchange)

• “culture wash”

• In-between samples for in-process

control (+ pictures) can be taken (at any

time-point of culture)

Minimal User Interaction• Attach starting product, media,

TransAct.

• Connection fresh virus (day 1)

• New medium bag (2L for 5-7 days)

• Connect solution for harvest (day 10-

14)



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T cell transduction and expansion process

Results

Cell

expansion

Cell

formulation

Cell

transduction

Cell

activation

Cell

selection

Final packaged

Cell product

CliniMACS

Reagents

MACS GMP

TransAct Kit

Transduction

with LV

MACS GMP

IL-2/7/15

TexMACS

Rebuffering

CliniMACS Prodigy

TS 520

Enrichment

• CD4/CD8 or CD62L

Activation

• 1x10E8 T cells

Transduction

Expansion

• Volume: 250 mL

• >2.0x10E9 T cells 1

Functional CAR T cells

• Cytotoxicity/ cytokine secretion

Cryopreserved

or fresh

apharesis

October 5, 2016

1) 3x106 CD19 CAR T cells/kg body weight (Davila et al. (2014) Sci Trans Med)CliniMACS Prodigy CAR-T Cells


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T cell cultivation – High cell densities and high viability

Cell

expansion

Cell

formulation

Cell

transduction

Cell

activation

Cell

selection

Final packaged

Cell product

Cryopreserved

or fresh

apharesis

0 2 4 6 8 10 12 14 16

0

5

10

15

0

100

200

300

time (in days)

Glu

cose (g/L

) pH

valu

e

volume

via

bility

(in %

) tota

l volu

me (in

mL) s

hakin

g s

peed (in

rpm

)

type 3

0 2 4 6 8 10 12 14

0

10

20

30

40

50

60

0

50

100

150

200

250

300

time (in days)

ce

ll d

en

sity ( x

10E

6 )

tota

l cell c

ou

nt ( x10E

8 )

T c

ell e

xp

an

sio

n (x fo

ld)

volume

sh

akin

g s

peed

(in rp

m)

tota

l cu

lture

Vo

lum

e (in

mL

)

typ

e 3



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T cell cultivation- robust and reproducible

Cell

expansion

Cell

formulation

Cell

transduction

Cell

activation

Cell

selection

Final packaged

Cell product

Cryopreserved

or fresh

apharesis

count

(day 10-14)

T cells CAR+ T cells

non-transduced 3.11E9 (±1.3) -

transduced 3.13E9 (±0.9) 1.3E9 (±0.3)0 2 4 6 8 10 12 140

10

20

30

40

50

time [days]

cell

count [x10

8]

lentiviral transduced

non-transduced

0 2 4 6 8 10 12 140

25

50

75

100

time [days]

via

bili

ty [%

]

lentiviral transduced

non-transduced

Robust expansion during non-

transduced (n=6) and lentiviral

transduced (n=10) runs

Good viability of each

manufactured cellular product

E.g. dose: 106 CAR+ T cells / kg

(130kg patient)



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New processes being added in…..


1. Retroviral transduction (2016/17)

2. Electroporation (2016/2017)


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Everything you need to make CAR-T cells from one

supplier


One Supplier

One Stop For Quality Paperwork

One Audit

One Purchase/Supply Agreement

One Partner


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Donor /

patient

Blood

LeukapheresisPatient

Cell

selection

Cell

transduction

Cell

formulation

Cell

activation

Cell

expansion

• End-to-End Solution

• Standardised, reproducible, transferable

• Easy commissioning, servicing and QA.

• No connection/transfer complications between

different manufacturers

• Automated

• Manufacture and QC

• Standardised, Reproducible, Transferable.

• Minimised personnel and training considerations.

• Closed-system

• Instantly scalable.



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Applicable to Centralised Manufacturing…



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…Or Localised/Point of Care Manufacturing



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October 5, 2016

1 Prodigy

How many products a month?

How many staff does that require?

How many clean rooms?

How many pieces of equipment?


• 3

• 1 - 2

• 1

• 1

Phase I/II


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October 5, 2016

100 Prodigy’s

How may products a month?





• 300

• 20

• 1

• 100

Roll Out


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October 5, 2016

100 Prodigy’s

How may products a YEAR?





• 3,600

• 20

• 1

• 100

Roll Out

elevator pitch: engineering car t cells on the...

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