BioInnovation Leaders Sum

mit

London, February 2015

Integrated approach to online process m

onitoring in Cell C

ulture applications

F. Wolpers

Ham

ilton Bonaduz A

G

The Heart of A

nalytics

2

Chapters

1. P

rocess Control C

hallanges

2. A

rc Sensors

3. C

ell Density S

ensors

4. S

ingle Use S

olutions

5. S

umm

ary

6. A

bout Ham

ilton

3

Fermentation – the traditional approach

Use N

utrients + Microbes à

Mix &

Wait = Ferm

entation

Is your application much m

ore complex?

4

Control is critical for ferm

entation

No direct control for cells available:

u C

ell status most critical

u M

ainly offline methods used

u D

irect measurem

ent needed u Labor intensive

u Final results available at end of process

u N

ot continuous control, just a sample rate

Most online m

easurements are insufficient:

u pH

and DO

u A

nalog signals are weak and unstable

u S

ensor lifetime and m

aintenance is critical u not necessarily connected to the cell status

5

Offline M

ethods are prone to error

Take into consideration: u S

ample rate frequency enough?

u M

issing important events?

u P

rocess results available only upto 24 h after sampling

u Too late to interfere

u Loss of tim

e and work in case of failure

u C

heck growth rate

u A

nalyze process deviation u R

isk of sterility breach (reactor and sample)

6

Arc S

ensors - integrated online process control

Arc - Intelligent P

rocess Sensors

Integrated sensor intelligence eliminates

the need for external transmitters

u Fully com

pensated, digital signal à

Improved robustness and reliability of

signal

u S

implified troubleshooting

and built-in diagnostics

u In-lab calibration

u R

educe setup costs

u E

asy to integrate

u Validation friendly:

Docum

entation of sensor use history

7

AR

C C

oncept Overview

Conventional Sensor Setup

nA, m

V analogue

4-20mA

AR

C Sensor Setup

Modbus, 4-20m

A

DIGITAL

8

Arc P

roducts: pH

Different applications require different pH

sensors ► E

asyFerm P

lus ► E

asyFerm B

io ► P

olilyte Plus

► pH

easy

9

Arc P

roducts: Optical D

O m

easurement

VisiFerm/VisiP

ro DO

: proven method for

measurem

ent of oxygen in process u S

imple m

aintenance with no electrolyte

u R

ugged sensor cap in four versions for different applications

u N

o interference with C

O2

u N

o oxygen consumption

u Integrated intelligence

u N

o polarization u O

utput signal: EC

S, 4-20m

A, M

odbus or H

AR

T (in VisiPro D

O)

u S

tandard 12 mm

format

10

Online M

onitoring of Viable Cell D

ensity (VC

D)

u C

apacitance based measurem

ent

u A

ccurate and precise method to determ

ine living cell concentrations

u D

irect correlation of the biomass w

ith process controlling param

eters like pH and D

O

u Linked to biovolum

e, intracellular conductivity, m

embrane capacitance and

cell size distribution

u The m

easuring line consists of capacitance sensor, pre-am

plifier and transmitter

11

Gaining direct inform

ation

By using capacitance based B

iomass S

ensors: u Viable cells react as capacitor in alternating electrical field

V ~

dead cells not polarized

viable cells polarized sensor-tip

Platinum

electrodes

12

Do not m

iss “hidden” events

Identify events: u E

vent just by online measurem

ent detected u N

o detection when counting total cells

u Im

pact on viable cells

time (h

)

050

100150

200250

permittivity (pF/cm)

0 1 2 3 4 5 6 7 8

viable cell count (c/ml)total cell count (c/ml)0.0

1.0e+6

2.0e+6

3.0e+6

4.0e+6

5.0e+6

6.0e+6

7.0e+6

8.0e+6

9.0e+6

1.0e+7

permittivity 2L

viable cell count (hema)

total cell count (hema)

error bars indicate +/- 1 S

D

Courtesy of R

oche

Decrease in capacitance

indicates a nutrient depletion

13

Investigate cell status

Clarify events:

u A

fter event detection, correlation with additional offline

measurem

ents for clarification u N

utrients and by-products u O

ptimal know

ledge on process needed u N

utrient depletion

Courtesy of R

oche

14

Increase Efficiency

Nutrient lim

itation is reduced u Trigger feeding pum

ps based on cell status u H

igher viability and elongated growth tim

es, lead to higher yield

time (h

)

5075

100125

permittivity (pF/cm)

2 3 4 5 6 7 8 9

glutamine concentration (mmol/L)glucose concentration (g/L)0 1 2 3 4 5

alanine concentration (mmol/L)2 3 4 5 6 7 8

permittivity

glucosealanineglutam

ine

= tim

e of feed addition

15

Increased Productivity

Optim

ization of harvesting time:

u C

ells stop producing at «hidden» event for unknown reason

u R

ealized by continuous control of cell status u P

rocess time w

as decreased by two days

u 10%

higher productivity

Courtesy of M

erck Serono

Permitivity (pF/cm)

16

pH, D

O, V

CD

and TCD

now com

bined !!

Integration of Arc S

ensors, by using the Arc W

ireless Converter

NEW

17

View and R

ecord Arc S

ensors

Integration of Arc S

ensors into Arc View

Controller

Dissolved O

xygen C

ell Density

Conductivity / pH

NEW

Easyferm

Bio (pH

)

18

Reliable (!) S

ingle Use S

olution for DO

& pH

Available 2015

u A

rc Module plugged on O

neFerm S

U pH

or DO

Electrode

u A

rc Wi W

ireless Adapter on top of A

rc Module

u S

ystem pow

ered by Pow

er Cable

u W

ireless comm

unication to PC

with W

ireless Converter or A

rc View

Handheld – soon based on B

luetooth 4.0 u D

ata logging with P

C and H

DM

software or A

rc View H

andheld u C

onfiguration, calibration, display with P

C and H

amilton G

MP

Package softw

are or Arc View

Handheld

Arc W

i

Arc

Module

SU

pH

Electrode

PC

with H

DM

software and

wireless converter

19

Single U

se Solution for C

ell Density M

onitoring The Future

<Prototype-Status>

u U

nder development

u H

amilton Proprietary IP and Patent

u U

ses the same technology as the

conventional Biom

ass Sensor

u D

isk can be welded in any bag

u P

rocess and Media com

patible m

aterials u Final design still to be determ

ined

20

Sum

mary

Intelligent Arc sensors do resolve problem

s of analog sensors u S

ignal Robustness

u Longer Lifetim

e

Intelligent Arc sensors do offer m

any new features and benefits

u Lab C

alibration u P

ermanent S

elf-Diagnostic

u S

ensor History

O

nline Cell D

ensity Monitoring for increased productivity and yields

u B

etter understanding of process dynamics

u O

ptimized feeding strategy, inoculation and harvesting tim

e u Full process control together w

ith pH, D

O (and C

onductivity or OR

P)

21

60 Years Ham

ilton: A G

lobal Com

pany

500 Em

ployees 800+ E

mployees

Reno, N

evada u 

Syringes

u 

Pipettes

u 

Diluters and D

ispensers u 

OE

M com

ponents u 

Robotic Liquid H

andling

Franklin, Massachusetts

u 

Autom

ated -20/-80�C

freezers

u 

Cryo Vial A

utomated

DeC

apper u 

Rack R

unner

100 Em

ployees

Timisoara, R

omania

u 

Syringes

u 

Lab-Trade Products

90 Em

ployees

Sales and S

ervice Organisations

Bonaduz, S

witzerland

u 

Syringes

u 

Lab and Process

Sensors

u 

Robotic Liquid H

andling u 

Medical Ventilators

22

Ham

ilton Bonaduz A

G

Via Crusch 8

CH

-7402 Bonaduz

Sw

itzerland Tel.: + 41 58 610 10 10 Fax: + 41 58 610 00 10

contact.pa.ch@

hamilton.ch

ww

w.ham

iltoncompany.com

Thank you for your attention