Designing Robust Virus Filtration Processes to Maximize ... · Virus filtration Virus filtration 2...
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Designing Robust Virus Filtration Processes to Maximize Operational Flexibility While Achieving Small Virus Removal
Daniel Strauss, PhDSenior ScientistAsahi Kasei Bioprocess [email protected]
BPI West 2017March 2nd, 2017San Francisco, CA
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Definition of Robustness
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� Property of being strong and healthy inconstitution
� For a system: ability of toleratingperturbations that might affect thesystem’s functional body
� For an object: sturdy in construction
� For a system: able to withstand or overcome “adverse conditions ”
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1) Robustness & Virus Removal
2) Robustness & Filtration Flux/Capacity
3) Robustness & Nanofilter Quality
Content
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Planova Nanofilters
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Planova TM 20N Planova TM BioEX
� Parvovirus removal nanofilters� Launched respectively in 2001 and 2009� Size exclusion mechanism and “multi-layer” filtration
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1) Robustness & Virus Removal
Content
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Robustness & Virus Removal
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� Goal: tolerating perturbations to keep LRV > 4 log
� “Perturbations” : concentration, pH, conductivity, clogging/fouling, process pause
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Robustness & Virus Removal
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Impact of Protein Concentration:
Tomoko Hongo-Hirasaki, Asahi Kasei Medical, 2011 PDA Conference
Virus safety assured even if change of conc. during processdevelopment (phase I/III) ➔ process optimizations possible !
� Increase of concentration ➔Decrease of flux
� Increase of mass throughput up to a certain point
� No impact on PPV LRV (> 4 log)
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Robustness & Virus Removal
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Impact of pH :
Virus safety assured even if change of pH during processdevelopment (phase I/III) ➔ process optimizations possible !
� Impact on flux
� No impact on PPV LRV (> 4 log)
Tomoko Hongo-Hirasaki, Asahi Kasei Medical, 2011 PDA Conference
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Robustness & Virus Removal
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Impact of Ionic Strength:
Tomoko Hongo-Hirasaki, Asahi Kasei Medical, 2011 PDA Conference
� Impact on flux
Virus safety assured even if change of cond. during processdevelopment (phase I/III) ➔ process optimizations possible !
� No impact on PPV LRV (> 4 log)
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Robustness & Virus Removal
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Impact of Clogging/Fouling on Planova TM BioEX:
Theory: smaller pores clog first➔ virus passage increases with clogging
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▲F
lux
deca
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)
●F
lux;
3%
h-I
gG (L
/m2/
hr)
Filtration time (hour)
0123456
0 20 40 60 80 100Flux decay (%)
PP
V L
RV
(fr
actio
n)
Virus safety assured even if unexpected production problemleading to big flux decay (nevertheless deviation must beaddressed…)
Koichiro Yanagida, Asahi Kasei Medical, 2009 Planova Workshop – adapted
No impact on PPV LRV (> 4 log)
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� Plasma-derived IgG on Planova 20N
� 20 hour process pause duration
Complete PPV clearance is achieved even at 20 h of pause
Pause20 h
Pre
ssur
e
0.8 bar100 L/m2
Virusfiltration
Bufferchase
0.8 bar30 L/m2
0 bar
IgG concentration: 10 mg/mL
Virus titer in theprotein solution:
6.2- 6.3 log10 TCID50/mL
Solution pH & NaCl Concentration
Robustness & Virus Removal
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� Plasma-derived IgG on Planova 20N
� Process pause at low pH, high ionic strength
• Pool LRV > 4 is maintained even at 20 h of pause
• Minimizing pause duration minimizes the impact
Pause
Pre
ssur
e
0.8 bar100 L/m2
Virusfiltration
Bufferchase
0.8 bar30 L/m2
0 bar
IgG concentration: 10 mg/mL
Virus titer in theprotein solution: 6.4 - 6.7 log10 TCID50/mL
Solution condition: pH 4, 100 mM NaCl
Robustness & Virus Removal
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Complete PPV clearance is observed under all condition s
Solution pH & NaCl Concentration
Pause3 h
Pre
ssur
e
2 bar100 L/m2
Virusfiltration
Virusfiltration
2 bar50 L/m2
0 bar
IgG concentration: 10 mg/mL
Virus titer in theprotein solution:
6.0- 6.4 log10 TCID50/mL
Robustness & Virus Removal� Plasma-derived IgG on Planova BioEX
� Process pause at low pH, high ionic strength
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The impact of process pause is dependent on the amo unt of virus challenge used during the spiking study
2015 Planova Workshop, Tomoko Hongo-Hirasaki, et al. (adapted)
Planova™ 20N filterProtein Concentration: 10 mg/mL h-IgGSolution Conditions: pH 7, 100 mM NaClVirus Spike: 5.1, 6.4, 7.0 log10TCID50/mL
Robustness & Virus Removal
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2) Robustness & Filtration Flux/Capacity
Content
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Robustness & Flux/Capacity
� Goal: tolerating perturbations to keep
• Small Flux decay or High Vmax
• Consistent Flux (L/m2/h) or Filtration Capacity (L/m2)
� Main “perturbations” : protein, concentration, virusspiking
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Robustness & Flux/Capacity
Impact of Protein:
Josh Goldstein, Janssen, 2014 Planova Workshop conference
� Similar levels of flux for the 3 mAbs� Consistent flux with no or small decay� Ideal for platform or multi-product plant� Same trend for PlanovaTM BioEX
Mabs with different pI tested with different pH & Conductivities
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Robustness & Flux/Capacity
Impact of Concentration:
� Impact of concentration on flux/capacity
� But only ~ 30% flux decrease while concentration x 2 for 10-20 g/L
Planova filters can be considered robust against concentra tionincrease up to a certain point
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h-IgG concentration (mg/mL)(100 mM NaCl, pH 4.5)
Filt
ratio
n C
apac
ity (
L/m
2 /3h
)
BioEX (294 kPa)
20N (78 kPa)
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Robustness & Flux/Capacity
Impact of Concentration:
� Optimal concentration to get the maximum mass throughput (kg or g /m2)
� Case by case !
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Robustness & Flux/Capacity
Impact of Concentration:
10 kg IgG nanofiltration:
� 10 mg/mL: - 20N: 1.5 kg/m2
➔ 6.7 m2
- BioEX: 3.5 kg/m2
➔ 2.9 m2
� 30 mg/mL: - 20N: 2.5 kg/m2
➔ 4 m2
- BioEX: 6.5 kg/m2
➔ 1.5 m2
Increase of protein mass throughput (g/m 2):➔ smaller filtration surface ➔ more cost effective
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Robustness & Flux/Capacity
Impact of Virus Spiking:
� Trend of most of the nanofilters on the market
� Oversizing of filtration surface area
� What about Planova filters?
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Robustness & Flux/Capacity
Impact of Virus Spiking:
20N
1 g/L h-IgG
50 mM Acetate, 100mM
NaCl, pH 5
Pressure: 14.2 psi
BioEX
3 g/L h-IgG
50 mM Acetate, 100mM NaCl,
pH 5
Pressure: 47 psi
Preparation of Ultra 1, 2, 3 and experiments conducted at Wuxi AppTecInc.
� No impact of virus spiking material on flux� Consistent flux with small decay� No oversizing of the nanofilter !
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3) Robustness & Nanofilter Quality
Content
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Robustness & Nanofilter Quality
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The last 2 important questions:
� Is the Robustness kept during Process Scale-up ?
� Is the Robustness kept from one Nanofilter Lot toanother nanofilter lot ?
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Robustness & Nanofilter Quality
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Brian Buesing, Asahi Kasei Bioprocess, 2015 Planova Workshop conference
20N and BioEX
3.40 bar (49.7 PSI)
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Robustness & Nanofilter Quality
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Robustness & Nanofilter Quality
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Brian Buesing, Asahi Kasei Bioprocess, 2015 Planova Workshop.
Consistent flux observed at all scales and no lot t o lot variability
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Robustness & Nanofilter Quality
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Brian Buesing, Asahi Kasei Bioprocess, 2015 Planova Workshop.
and lots
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Robustness & Nanofilter Quality
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Daniel Strauss, Asahi Kasei Bioprocess, 2016 Planova Workshop.
Consistent flux observed at all scales and no lot t o lot variability
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Robustness & Nanofilter Quality
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Daniel Strauss, Asahi Kasei Bioprocess, 2016 Planova Workshop.
Robust Viral Clearance across all filter sizes and l ots
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1) Process Development:� Possibility to change conditions (buffer, concentration, pH, cond.)
to optimize nanofiltration or other purification steps
� Or faster development w/o optimizations because “it simply works”
2) Virus Clearance Validation:� High LRV achievable
� No oversizing of the nanofilter
3) Manufacturing & Commercial Production:� Process pause manageable
� Virus safety ensured despite accidents or purification problems
� Multi-product platform approach
� Slight production deviations absorbable
� Reliable performances
Conclusion - Concrete Benefits
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Acknowledgements
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Thank you to my colleagues in Japan, US and Europe !
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どうもありがとう
Domo Arigato!
(thank you very much)
Questions ??