applying qbd to biotech process validation
DESCRIPTION
In this session from the Institute of Validation Technology's Validation Week Europe, Kurtis Epp and John Kandl discuss how to implement QbD to all three stages of process validation.TRANSCRIPT
BIBioTech
BIApplyingValidationA case stuStage 1 –
TECHLOGIC
BioTechLogic, Inc.BIO TECH
LOGICBIO®
TECHIO LOGICLogic, Inc.
IO®
g QbD to Biotech Process n :udy in applying QbD to
– Process Design
IVT Validation Week 28-30 March 201128 30 March 2011
Kurtis Epp, John KandlBioTechLogic, Inc.
Agenda
• Process Backgrou• Risk Assessment• DOE• Parameter Evaluat• Conclusions
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
und
tion
Slide 2Company Confidential
Process Back
The scope of this case stupdata obtained from small-Chromatography runs perlimits in experiments planlimits in experiments planDesign of Experiments (D
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
kground
udy is to provide and evaluate y p-scale Ion Exchange rformed within defined parameter ned and executed according toned and executed according to
DOE).
Slide 3Company Confidential
Process Back
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
kground
Slide 4Company Confidential
Risk AssessmOperating Parameter Normal Operating
UV at Start of AEX Fraction Collection > 0.50 AUUV at End of AEX Fraction Collection < 20% of maximum p
Elution Gradient 8% per CVElution Gradient (20 – 100% B oveEquilibration volume ≥ 5 CVLoad flow rate 100 – 120 L/h (TargeWash flow rate 100 – 120 L/h (TargeElution flow rate 100 – 120 L/h (TargePre-Equilibration flow rate 100 – 120 L/h (Targe
F1 F3: 0 2Fraction Volume
F1 – F3: 0.2 F4+: 0.1 CV
Wash volume ≥2 CVFraction Mixing Speed 65 rpmFraction Mixing Time 20 - 30 min
Pool Mixing Time 10 – 15 minPool Mixing Time
Pool Mixing Speed 100 rpm
Pre-Equilibration volume ≥ 4 CVWFI Rinse volume ≥ 3 CVWFI Rinse flow rate 100 – 120 L/h (TargeEquilibration flow rate 100 – 120 L/h (Targeq ( g
In-Process Control LimitsColumn Load 15 – 25 g / L rColumn Bed Height 30 ± 3 cmColumn Backpressure during Equilibration, Load, Wash, and Elution < 3 bar
Effl t H t d f E ilib ti ± 0.3 pH Units of EqEffluent pH at end of Equilibration p qBuffer pH
Effluent Cond. at end of Equilibration ± 1 mS/cm Units of EBuffer Conduc
Effluent UV at the end of Equilibration ZeroFraction Pooling Criteria (RP-HPLC) ≥ 95% Main P
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
mentg Range Potential Effect(s) of Failure SEV OCC DET RPN
U Product loss, quality 9 4 2 72peak height Product loss, quality 9 4 2 72V Inconsistent quality 8 2 4 64r 10 CV) Inconsistent quality 8 2 4 64
Product loss, quality 9 2 1 18et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Longer equilibration 4 1 3 12CVCVV Product loss, quality 9 1 1 9
Inconsistent quality 5 1 1 5Inconsistent quality 4 1 1 4
n Inconsistent quality 4 1 1 4
n Non-homogeneity, inconsistent sampling/yield 4 1 1 4sampling/yieldNon-homogeneity, inconsistent sampling/yield 4 1 1 4
Longer equilibration 3 1 1 3Longer equilibration 3 1 1 3
et: 110 L/h) Longer equilibration 1 1 1 3et: 110 L/h) Product loss, quality 3 1 1 3) , q y
resinm
quilibration q
Equilibration ctivity
Peak
Slide 5Company Confidential
Risk Assessm
The parameters selectedpthe UV at start of fractioncollection, gradient slopeD d t i blDependent variables anaand purity by RP-HPLC.
TestinInput parameter
Lower
UV at the beginning of fraction collection (AU) 0.40
UV at the end of pooling (% from peak max) 10
Gradient slope (% per CV) 4.0
Resin Load, (g/L resin) 15
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
ment
d for evaluation in this study were yn collection, UV at end of fraction e, and resin load.
l d t i i ldalyzed were protein recovery yield
ng limitsgOutput parameters
Upper
0.60
Purity by RP-HPLC (≥ 95%)Step yield (≥ 75%)
30
12.0
25
Slide 6Company Confidential
DOE
Pattern Exp. No.:
UV at the staof fractioncollectionp collection
(AU)−−−− 1 0.40000 2 0.50000 3 0 50000 3 0.5−++− 4 0.40000 5 0.5++−− 6 0.6
7 0 6+−−+ 7 0.6++++ 8 0.6+−+− 9 0.6−−++ 10 0.40000 11 0.5−+−+ 12 0.4
Note: It is important to randomizeindependence of your observationmistake by the operator.
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
art n UV at the end
of pooling Gradient
slope Resin Load (g/L)(% peak max) (% per CV) (g/L)
10 4 1520 8 2020 8 2020 8 2030 12 1520 8 2030 4 1510 4 2510 4 2530 12 2510 12 1510 12 2520 8 2030 4 25
e the run order to assure the ns and reduce the chances of a
Slide 7Company Confidential
Experimental
• A summary of the varIon Exchange ChromOutputs step yield anOutputs, step yield anlisted in the following
• Runs that did not meeare shaded to indicate
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
Results
riable parameters for each of the atography runs as well as the
nd eluate purity by RP HPLC arend eluate purity by RP-HPLC are table.
et the defined acceptance criteria pe run failure
Slide 8Company Confidential
Experimental
Run Resin Load GradientRun Number
Resin Load (g/L)
Gradient Slope (%)
1 15 4
2 20 8
3 20 8
4 15 12
5 20 85 20 8
6 15 4
7 25 4
8 25 128 25 12
9 15 12
10 25 12
11 20 811 20 8
12 25 4
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
Results
UV @ Start % Peak Step Yield (%)
RP-HPLC Purity (%)UV @ Start
Collection Height End Collection
(%) Purity (%) Acceptance Criteria≥ 45% ≥ 95%
0.4 10 53 95
0.5 20 56 97
0.5 20 50 97
0.4 30 59 96
0 5 20 51 970.5 20 51 97
0.6 30 53 97
0.6 10 52 96
0 6 30 40 970.6 30 40 97
0.6 10 50 98
0.4 10 45 97
0.5 20 51 970.5 20 51 97
0.4 30 60 97
Slide 9Company Confidential
Statistical dat
• For evaluation of statistion protein quality and qsoftware package was uSquares model for EffecSquares model for Effec
• The model was run sepof yield and purity by RP
• The fractional factorial minclusion of all single facinteractions as model efinteractions as model efresource and time constwas not possible.
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
ta analysis
ically significant effects of factors uantity, the JMP statistical used applying the Standard Least ct Leveragect Leverage. arately for two output parameters P-HPLC.model was initially run with the ctor and some two-factor ffects (Resolution IV) Due toffects (Resolution IV). Due to traints, a higher resolution study
Slide 10Company Confidential
Statistical Model PStep Yieldp
• The final model used for effects and two factor int
• There were two significai ifi t i t ti Tsignificant interaction. T
graphically represented iplots and interaction conp
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
Parameter Estimates for
assessment all four primary p yteractions listed above. nt main effects and one h l ihese conclusions were in the main effects leverage tour plots.p
Slide 11Company Confidential
Main Effect Lever
a. UV at Start Collection
c. Gradient Slope
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
rage Plots for Step Yield
b. UV at End Collection
d. Resin Load
Slide 12Company Confidential
Statistical Model PPurity by RP-HPLy y
• The final model used forprimary effects (excludintwo-factor interactionstwo factor interactions.
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
Parameter Estimates for C
r assessment included all four ng resin load) as well as three
Slide 13Company Confidential
Two-Factor InteraPurity by RP-HPLy y
Step Yield (%)
Gradient Slope (%/CV)
0
UV @ End (%)
a. UV at End Collection x Gradient Slope
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
action Contour Plots for C
Step Yield (%)
Resin Load (g/L resin)
0
UV @ Start (AU)
b. UV at Start Collection x Gradient Slope
Slide 14Company Confidential
Confirmation • There is one area of th
represents process failp pyield, run 8. In order toacceptance criterion isoperated within its defioperated within its defimodified and two additaugment the initial des
• We chose to tighten thas it was determined tosignificant primary effesignificant primary effeanalysis.
• None of the experimenpfailed the acceptance cconfirms the predicted study
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
study.
of Resultshe tested design space that lure space with regard to step p g po ensure that the step yield always met when the process is ned PARs the design space wasned PARs, the design space was tional runs were performed to sign.e gradient slope upper limit to 11% o be easy to control and the most ect for step yield in our initialect for step yield in our initial
ntal runs for the tightened limits gcriterion for step yield. This design space from the initial
Slide 15Company Confidential
Design SpaceSpaceSpace
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
e vs Defined Control
Based on the models generated for h f th d fi deach of the defined process
outputs, a three-dimensional plot was generated to graphically show the process control space with
d h i llregard to the experimentally defined design space
Slide 16Company Confidential
Conclusions
• The results of this DOdesign space from the
• Based on our findings h f th thwe chose for the three
appropriate with the ex• As a result we tighteneAs a result we tightene• The IPC limits for resin
set appropriately.
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
E study confirm the predicted y pe initial study.
we determined that the IPC limits i t te input parameters were
xception of Gradient Slope. ed our control limitsed our control limits.n load were also determined to be
Slide 17Company Confidential
Final Paramet
Normal OpeOperating Parameter
Normal Ope(NO
UV at the beginning of pooling, % from peak max Target: >p g p
UV at the end of pooling, % from peak max Targe
Gradient slope, % per CV TargeGradient slope, % per CV Targe
Performance Parameter In
Resin Load, g/L
* Note: The claimed experimentally confirmed range is slightly tighter than thsupport the higher end of the range.
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
ter Limits
rating Range Experimentally Confirmedrating Range OR)
Experimentally Confirmed Ranges
> 0.5 AU 0.4 – 0.6 AU
t: 20% 10 – 30%
et: 8% 4 – 11%*et: 8% 4 11%n-Process Acceptance Criterion
15 – 25 g/L
he design space in that it takes the worst-case data point to
Slide 18Company Confidential
Thank You
BioTechLogic Inc serBioTechLogic Inc. serbiopharmaceutical indwith companies to mwith companies to mresource and comme
TECHLOGICBIO TECHLOGICBIO
®BioTechLogic, Inc.
rves therves the dustry by collaborating eet their developmenteet their development,
ercialization needs.
www.biotechlogic.comwww.processvalidation.com
Slide 19Company Confidential