Sartorius - November 2021 - Simplifying Adeno Associated Virus - 27

MVDA
Initial Conditions
The DOE was used to identify optimum
operating conditions. To enhance the
analysis MVDA was used, this can be
helpful to increase process understanding
or to support trouble shooting.
Typically, this incorporates data
collected over time corresponding to
the process evolution, from the start
to the end of the batch using online
sensors (such as pH and DO), process
parameters (such as oxygen flow) or
at-line analytics (such as viable cell
count or metabolites). MVDA compresses
data in a way the batches can
be compared quickly for outliers,
trends, or clusters (Figure 4).
MVDA Analysis
In this second part, the additional process
information generated in the
DOE study was condensed into one
point, using MVDA, to compare all processes
in order to spot trends, groups,
or outliers. In this case a hierarchical
modeling concept was used for merging
DOE data with time series data.
EvolutionOutputs
Variables
1.5
1
DOE
0.5
-0.5
-1
-1.5
-1
-0.5
00.5
1
Figure 4: Combining DOE and MVDA data of Sf9 for rAAV production
The output from the hierarchal modeling
is seen in Figure 5, where each batch's
evolution trajectory has been condensed
into one single dot on the score scatter
plot. The plot clearly shows that one dot
represented CS2_V4 Batch is an outlying
event. This is also one of the batches with
the highest titer, as seen by the colouring
in the plot below.
A major advantage of MVDA is the possibility
to drill down to explain differences
observed. In this case the evolution data
was different for CS2_V4-Batch compared
to the average run (indicated by the yellow
bar in Figure 6).
Drilling further down links that to individual
process parameters (Figure 7) which
shows that the CS2_V4-Batch required
more oxygen. This is thought to be related
to higher cell growth and more cells can
lead to higher titer (assuming similar specific
productivity).
Although this cannot be immediately
changed, it is important to note and
understand this observation when
considering a scale-up with respect to
oxygen supply.
Figure 5: Summary of trajectory data from all batches of Sf9 cells transiently producing rAAV8 in Ambr®
15 micro bioreactors.
Titer 96 h
4
3
2
1
-1
-2
-3
-4
-4
-3
27 | GENengnews.com
-2
1e+12
CS2_V10-Batch
CS2_V9-Batch
CS2_V12-Batch
8e+11
CS2_V2-Batch
CS1_V4-Batch
CS2_V12-Batch
CS2_V7-Batch
CS2_V8-Batch
CS1_V1-Batch
CS1_V11-Batch
CS2_V1-Batch
CS2_V3-Batch
CS1_V2-Batch
CS1_V3-Batch
CS2_V6-Batch
CS1_V10-Batch
CS1_V8-Batch
CS2_V5-Batch
CS1_V7-Batch
CS1_V6-Batch
CS1_V9-Batch
2e+11
CS2_V4-Batch
4e+11
6e+11
-1
01
t[1] × 1.000007
23 45 6
Missing
to(1) × 1.25957
Time
Batches

Sartorius - November 2021 - Simplifying Adeno Associated Virus

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