MBI F4 Fermentation Process Operation Sensors, Monitoring and ...

An introduction to a novel
filtration system


ATF
-
manufacturing Platform

Upstream Unit Operations


ATF
-
cellcultivation

Concentrated Perfusion

Concentrated Fed
-
Batch

Hybrid Processes




2

Capacity cannot be calculated solely by bioreactor size

10kg

500g

1000L

3kg

150g

300L

1kg

50g

100L

20 Runs/Yr

Per Run
*

Fed
-
Batch

150kg

15kg

500g

1000L

45kg

4.5kg

150g

300L

15kg

1.5kg

50g

100L

10 Runs/Yr

30 Day Run

Per Day
*

Perfusion

* 500 mg/L post purification yield

15x more product

in the same time!

Avid Bioservices Facilities

Why use Perfusion in Research, even if not later...

•
Perfusion produces more material in a shorter time period

–
Do you have time to make a new cell line?

–
Do you have time to develop a special media?

•
Pre
-
clinical material can be produced faster with less development effort

•
A low producing cell line can be used to produce enough material in a
shorter time for a phase 1 trial

•
Smaller bioreactors are required at all stages of PD and Manufacturing
-

saving significant capital cost

•
One continuous culture experiment can perform multiple metabolic studies to
understand clone behavior in different design spaces, significantly faster
than in batch or fed
-
batch

•
Upstream development times may be reduced due to simpler feeding

Perfusion and Concentrated Perfusion

•
Perfusion is
dead

-

Long Live
Concentrated

Perfusion !!


•
What

is
Concentrated

Perfusion?

–
Only

possible

with the ATF

–
Ultra
-
high

viable

cell
densities
, in the region of
60
-
150m
/ ml

–
Requires

1
-
3
vv

/
day

media (not
more
)

–
Same
simple process
control

as
perfusion

& CFB
–

but

higher

productivity


•
When

should

perfusion

be
used

and not
Concentrated

Perfusion?

–
When

the
product

quality

is
affected

by cell
concentration
, or
if

protein
expression
depends

on a
dilute

toxic

species


Concentrated
Perfusion

•
“Standard” perfusion application but with the ATF System

–
high cell concentration
(60
-
100m/ml), 30
-
60 days

•
Straight
-
forward

feeding

control
,

removal of
metabolites

& proteins

•
Continuous

filtered

harvests
,
often

pooled

(
every

3
-
6
days
) for
easier

DSP

–
~0.1g/L/day per 10m cells /ml for an average cell line (10pg/cell/day at 2vvd)


M
F
filter
retains
cells inside
reactor while
molecules filter
through

Filtrate pump runs
at 0.5
-
3 vv / day

or as required by
cell growth

ATF rate constant
at minimum

~0.05
-
0.1 vv /
min

Feeding
controlled by
level probe

Graph taken from Genetic Engineering News October 2007

CHO Perfusion Results