bils 2015, f. wolpers hamilton bonaduz ag
TRANSCRIPT
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