Sartorius - November 2021 - Simplifying Adeno Associated Virus - 22

According to Zuban different statistical
metrics are used to assess model quality.
Two metrics of importance are R2
(model fit) and Q2 (predictive capability).
Higher numbers result in a better model
indicating that data is reproducible and
therefore the model is predictive.
One example of the data visualization
generated using a DOE approach is a
coefficient plot. This shows which factors
influence the response (in this case rAAV
titer). For example, using the coefficient
plot in Figure 1, larger positive bars on
the plot are seen with the cell line 1 and
transfection reagent 1 indicating that
these factors have a positive influence
on the titer.
The factors that are most crucial are then
carried over to an optimization design for
a more thorough investigation in the
next study. Zuban says: " Using DOE for
rAAV process optimization, you start
broad, for example, assessing the impact
from multiple factors. The derived model
is then challenged based on knowledge
and experience by the bioprocess scientists.
Using MODDE®
software for example,
the coefficient plots generated are easy
Observed vs. Predicted - Titer qPCR
11
10.5
10
9.5
9
1
8.5
8
11 17
7.5
7
6.5
6.5
77 8
.5
8.5
9
Predicted [vg/mL]
N=24, DF=17, Cond. no.=3.139, RSD=0.328, Q2=0.797
Figure 2: Observed-vs-predicted plot (left) and summary of fit plot for the model (right) for rAAV2/8 plasmids vector
and helper rAAV titer obtained from transiently transfected HEK293 cultured in Ambr®
15 micro bioreactors
22 | GENengnews.com
9.5
10
10.5
11
Titer qPCR
Spec. prod.
12
18
R2
Q2
Model validity
Reproducibility
8
2
3
19 16
24
23 15
9
5
10
14
21
4
6
7 22
20
13
Coefficients (scaled and centered) - Titer qPCR (vg/mL)
0.8
0.6
0.4
0.2
-0.2
-0.4
-0.6
-0.8
Cell line 1Cell line 2 Medium 2 Medium 1Cell
Density
Transfection
Reagent 1
Transfection
Reagent 2
N=24, DF=16, Cond. no.=1.414, RSD=0.541, Q2=0.031
Figure 1: Coefficients plot of the rAAV viral genome titer of a transient rAAV2/8 plasmids vector and
helper production with two HEK293 cell lines cultured in different media in Ambr®
to understand and they are a convenient
way of visualizing if a factor has a negative
or positive effect on process outcome.
From there you can choose the
most influential factors for the next DOE
Plasmid
ratio
Ratio Total plasmid
15 micro bioreactors
and using other published data and inhouse
knowledge on rAAV in suspension
cell culture to narrow down the right
ranges to test. "
Observed [vg/mL]
Titer [vg/mL]
https://www.genengnews.com/

Sartorius - November 2021 - Simplifying Adeno Associated Virus

Table of Contents for the Digital Edition of Sartorius - November 2021 - Simplifying Adeno Associated Virus

Sartorius - November 2021 - Simplifying Adeno Associated Virus - 1
Sartorius - November 2021 - Simplifying Adeno Associated Virus - 2
Sartorius - November 2021 - Simplifying Adeno Associated Virus - 3
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 5
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 8
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 11
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 15
Sartorius - November 2021 - Simplifying Adeno Associated Virus - 16
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 18
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 24
Sartorius - November 2021 - Simplifying Adeno Associated Virus - 25
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 27
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Sartorius - November 2021 - Simplifying Adeno Associated Virus - 31
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