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Model Based Scale up of Depth Filtration for Clarification of
Monoclonal Antibodies (mAb) Cell Culture Harvest
Vishwanath S Hebbi1, Sandeep Hadpe1, Muthukumar Sampath1, Souhardhya Roy1, Anurag S Rathore1,*
1Indian Institute of Technology Delhi, New Delhi, India
* Corresponding author. Tel: +91-11-26591098, Fax: +91-11-26581120, E-mail: [email protected]
Depth filtration is generally used post centrifugation for reducing Normalized
Turbidity Units (NTU), HCP and HCDNA levels. The operation of depth filtration
and its scale up is straightforward. However, due to various issues such as
fouling and variability in feed, 15-20% of area is considered in excess of
calculated area during the scale up. Therefore it is important to know the
plugging behavior of depth filters and their effect on scale up.
The present work targets this issue by proposing a model based scale up
wherein the different fouling mechanisms are investigated for constant flow
depth filtration. Interestingly, amongst the various models investigated,
combined model explains the fouling mechanism and the corresponding
parameters of the same model are used for scale up. The model based scale
up provides insights in deciding the accurate filter area during scale up.
Abstract
Methodology
Filtrate from Primary filter
D0HC 30SP B1HC 90ZA
Cell culture
Secondary FiltrationPrimary Filtration
Empirical modelling
Model based scale up
Validation scale up trails
Empirical modelling-Primary filtration
Observations
0
2
4
6
8
10
12
14
0 250 500 750 1000 1250 1500 1750
P/P
0
Time, t (s)
D0HC
Experimental Cake Intermediate
Complete Standard Cake Complete
Cake Intermediate Complete Standrd Intermediate Standard
Cake Standard
0
5
10
15
20
25
0 250 500 750 1000 1250 1500
P/P
0
Time, t (s)
30SP
Figure 1. Modelling of primary filtration for D0HC and 30SP
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 1.19E+06 13.00 1.62
Intermediate 22.43 3.93 0.89
Complete 5.18E-04 79.70 3.99
Standard 11.41 39.02 2.79
Cake Complete 2.44E-04 5.67E+05 1.58 0.63
Cake Intermediate 3.18E+05 7.56 1.01 0.50
Complete Standard 1.00E-04 11.60 32.06 2.83
Intermediate Standard 22.44 0.00 4.99 1.12
Cake Standard 4.79E+05 11.62 2.81 0.96
Table1. Model parameters of primary filtration for D0HC
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 2.98E+06 53.68 3.28
Intermediate 31.62 51.56 3.21
Complete 5.36E-04 510.50 10.10
Standard 13.63 311.90 8.83
Cake Complete 2.38E-04 1.49E+06 5.57 1.36
Cake Intermediate 9.74E+05 7.41 3.34 1.06
Complete Standard 1.00E-04 14.29 270.00 8.22
Intermediate Standard 21.50 0.91 220.80 7.43
Cake Standard 1.61E+06 14.12 13.65 1.85
Table2. Model parameters of primary filtration for 30SP
Empirical modelling-Secondary filtration
0
5
10
15
20
0 250 500 750 1000 1250 1500
P/P
0
Time, t (s)
B1HC
Experimental Cake Intermediate
Complete Standard Cake Complete
Cake Intermediate Complete Standrd Intermediate Standard
Cake Standard
0
5
10
15
20
25
0 400 800 1200 1600 2000 2400
P/P
0
Time, t (s)
90ZA
Table 3. Model parameters of secondary filtration for B1HC
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 8.16E+06 13.67 1.65
Intermediate 18.37 0.24 0.22
Complete 5.03E-04 17.84 1.89
Standard 10.85 4.78 0.98
Cake Complete 3.38E-04 2.27E+05 0.29 0.31
Cake Intermediate 8.85E+04 10.59 0.23 0.28
Complete Standard 1.00E-04 10.36 3.37 0.82
Intermediate Standard 17.52 0.24 0.21 0.23
Cake Standard 1.23E+05 11.23 0.46 0.39
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 7.74E+05 8.28 1.09
Intermediate 15.80 1.05 0.39
Complete 3.74E-04 43.21 2.49
Standard 8.90 16.93 1.56
Cake Complete 1.90E-04 3.45E+05 0.41 0.29
Cake Intermediate 2.09E+05 5.81 0.25 0.22
Complete Standard 1.00E-04 8.12 13.87 1.41
Intermediate Standard 15.66 0.00 1.34 0.47
Cake Standard 2.90E+05 8.64 0.50 0.29
Scale up strategy
Filter
Small Large
Volume (L) area (m2) Volume (L) area (m2)
30SP 0.35L 270L 270L 1.9
90ZA 0.40L 350L 350L 1.4
Figure 2. Modelling of Secondary filtration for B1HC and 90ZA
Table 4. Model parameters of secondary filtration for 90ZA
Scale up validation
0
2
4
6
8
10
12
14
16
0 2000 4000 6000
P/P
0
Time, t (s)
30SP
Experimental Cake Intermediate Complete
Standard Cake Complete Cake Intermediate Complete Standrd
Intermediate Standard Cake Standard
0
5
10
15
20
25
30
0 2000 4000 6000 8000
P/P
0
Time, t (s)
90ZA
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 1.59E+06 34.15 1.76
Intermediate 14.69 0.70 0.25
Complete 1.77E-04 34.99 1.78
Standard 8.58 6.72 0.78
Cake Complete 1.27E-04 3.56E+05 0.86 0.29
Cake Intermediate 4.28E+04 11.62 0.95 0.31
Complete Standard 1.00E-05 8.30 8.82 0.90
Intermediate Standard 12.70 0.57 0.87 0.29
Cake Standard 2.86E+05 8.06 0.80 0.28
ModelPlugging parameter Error
Kb Kc Ki Ks SSE RMSE
Cake 2.88E+06 45.72 2.32
Intermediate 15.53 196.00 4.43
Complete 1.47E-04 10.35 1.71
Standard 7.50 602.00 8.18
Cake Complete 4.95E-05 1.80E+06 13.68 1.23
Cake Intermediate 1.99E+06 1.40 6.96 0.88
Complete Standard 1.00E-04 4.43 679.20 8.24
Intermediate Standard 1.46 5.65 792.20 9.38
Cake Standard 2.12E+06 6.43 24.99 1.67
• 30SP and 90ZA are found to be the best filters for clarification of cell culture
broth
• Cake intermediate model fits the experimental data with least error for both
primary as well as secondary filtraions
• Plugging behavior changed for primart filtration at 1.6 m2 scale
NTU Filtrate (ml)
D0HC 354 300
30SP 130 350
NTU Filtrate (ml)
B1HC 3.8 350
90ZA 1.8 350
Conclusion
• Depth filters are useful in clarification od mammalian cell culture broth
• Combined models demonstrates the plugging mechanism in depth filtrations
• Model parameters can be useful in sizing of depth filters
Future work
• Incorporation of flow behavior terms into scale up strategy at larger scale
filtration
• Generalization of scale up criteria for depth filtration
• To study the effect of other variables such as cell viability, NTU variation in the
load on plugging mechanism
Table 1. Performance of primary and secondary filters
References
1. Yavorsky, David, et al. "The clarification of bioreactor cell cultures for
biopharmaceuticals." Pharmaceutical technology 27.3 (2003): 62-77.
2. Ho, WS Winston, and Kamalesh K. Sirkar, eds. Membrane handbook.
Springer Science & Business Media, 1992.
Figure 3. Scheme for scale up for depth ffiltration
Time versus pressure data is generated for filter
Determination of plugging parameters
Vsmall and Vlarge ,Time
Assume area Alarge and fix Pmax
Calculate P from model equation
If
pcalc=Pmax
Yes
Assumed area is the final area
Assume diff value for Alarge
No
Table 5. Scale up result
Figure 4. Validation of scale up for 30SP and 90ZA
Table 6. Model parameters for scale up run for 30SP
Table 7. Model parameters for scale up run for 90ZA