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Validation in Biotech Facilities: What ? Why ? How?
Dr. PK Yegneswaran
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Presentation Outline
� Validation Overview
� Cleaning Validation
� Process Validation
� Sterilization Validation
� Citation Examples
� Regulatory References
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Typical Project Schedule
1 2 3 4 5 6
Scope
Design
Procurement
Construction
IQ/OQ
Startup / Validation
100%
YEAR
APPROVAL
% S
pe
nt
Phase III
APPROVAL
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Typical Post OQ Schedule
Cleaning Dev.
Sterilization Dev.
Practice Lots
IQ/OQ/Facility/Utility Qualification
Validation Lots
YEAR
APPROVAL
Phase III
APPROVAL
1H Y4 2H Y4 1H Y5 2H Y5
Cleaning Valdn.
Sterilization Valdn.
File Licens
e
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Validation Overview
[To establish] documented evidence which provides a
high degree of assurance that a specific process
will consistently produce a product meeting pre-determined specifications and quality attributes.
(FDA, May 1987)
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Validation Overview
� Why validate ?
– The FDA requires that we validate all of our
systems and processes according to 21 CFR part 211
– Improves our understanding of our manufacturing
processes
– Right thing to do !
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Why Validate?
� Consistent yield &
quality
� Rapid decisions when
mishaps occur
� Fewer discards
� Less time hosting government agencies,
more time
manufacturing
= to ensure that the
output is consistent;
first time, every time!!
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What do we Validate ?
� Process
� Cleaning
� Sterilization
� Filters
� Containers
� Assays
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How do we Validate ? Cleaning, Process, Sterilization etc.
� Define Critical Process Parameters, Critical Quality Attributes
� Develop protocol describing validation studies
– Consider fractional study approach for cleaning, sterilization
� Execute studies
� Address deviations
� Compile report
� Review / Approve report
� GMP Documentation all the way=.
This process applies to all validation
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Definitions
Critical Process Parameter (CPP):
An input variable that must be controlled within a specified range to ensure success.
Critical Quality Attribute (CQA):
An output parameter from a unit operation that must be within a specified range to demonstrate control, consistency, and acceptable product quality.
CPP CQA
Ionic Strength Ion Exchange Yield Column Load Chromatography Purity
Flow rate Cleaning Conductivity
Temperature TOC
Concn.
Sat. Steam Sterilization BIs
Time
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Style-ogen® Facility Built
IQ/OQ Complete What Next ?
• Sterilization Validation • Development, validation studies
• Practice Lots • Define validation parameters for process, cleaning, cleaning
validation
• Validation Lots • Process validation, cleaning validation
• File license
• Pre-Approval Inspection
• Facility / Product approval
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Typical Post OQ Schedule
Cleaning Dev.
Sterilization Dev.
Practice Lots
IQ/OQ/Facility/Utility Qualification
Validation Lots
YEAR
APPROVAL
Phase III
APPROVAL
1H Y4 2H Y4 1H Y5 2H Y5
Cleaning Valdn.
Sterilization Valdn.
File Licens
e
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Presentation Outline
� Validation Overview
� Cleaning Validation
� Process Validation
� Sterilization Validation
� Citation Examples
� Regulatory References
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Cleaning Validation
� Cleaning Validation overview
� Cycle development for Style-ogen®
equipment
� Validation of cleaning cycles
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Equipment cleaning validation is the process
of establishing documented evidence that a
particular cleaning procedure will consistently
reduce equipment surface residuals to a predetermined acceptable level.
“Residuals” are any product, degradate,
intermediate, excipient, raw material/reactant
or cleaning agent that may reside on any equipment surface following processing
and/or cleaning.
What is Cleaning Validation ?
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21 CFR Part 211 Subpart D - Equipment
211.67 Equipment Cleaning and Maintenance
– (a) “Equipment and utensils shall be cleaned,
maintained, and sanitized at appropriate intervals to prevent malfunctions or contamination that would alter
the safety, identity, strength, quality, or purity of the
drug productA.”
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21 CFR Part 211 Subpart F - Production and Process Controls
211.113 Control of Microbiological
Contamination
– (a) “Appropriate written procedures, designed to prevent
objectionable microorganisms in drug products not
required to be sterile, shall be established and
followed.”
– (b) “Appropriate written procedures, designed to prevent
microbiological contamination of drug products
purporting to be sterile, shall be established and
followed.”
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One-Third Of Recent Drug GMP Warning Letters
Cite Cleaning Practices (2002 survey)
Year # of Warning Letters
# of Warning Letters related
to Cleaning
% Related to Cleaning
1999 65 23 35%
2000 71 20 41%
2001 71 20 28%
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Cleaning Validation Where Do I Start?
Validation Strategy
Manufacturing
Process
Cleaning
Process
Mfg. Equipment
& Design
Manufacturing
Process
Cleaning
Process
Mfg. Equipment
& Design
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� Obtain a Process Flow Diagram
� Is Product Inactivation Required ? (Important to ensure inactivation procedure is defined before starting validation)
� Define Applicable Hold Times � Dirty: End of Process to Start of Cleaning � Clean: End of Cleaning to Next Process Use � Sterile: End of SS/SIP to Next Process Use
� What “Residuals” Need to Be Cleaned by the CIP ? � Product (includes degradates, excipients, raw materials, etc.) � Cleaning Agents
� Are the “Residuals” Representative of the Process ? (Important to consider when validating during Practice Runs or Demonstration)
� Is the Equipt. Sanitized or Sterilized after CIP ?
Considerations:
Manufacturing Process
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� Define CIP Type (Manual, Automated)
� Are the Individual Steps of the CIP Procedure Defined ?
� Critical Process Parameters Defined ? (e.g. Flow, Temp.)
� Is the Cleaning SOP available ?
� Does CIP Procedure Clean All Product Contact Surfaces?
(Highlight and Compare Mfg Process to CIP Process on Same P&ID)
� What CIP Cycle Development Work is Planned?
Considerations:
Cleaning Process
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� Are All Equipt/Systems IQ/OQ’d ?
� Define Surface Materials of Construction (Prod. Contact)
� Ensure General CIP Design Principles Followed
� Minimal to No System Deadlegs
� Turbulent Flow Maintained During CIP
� Full Coverage to Vessel During the CIP
� Lines Flooded Completely During the CIP
� Complete System Drainability
� Assess Validation Sample Locations
� Accessibility
� Availability
Considerations:
Mfg. Equipment Design
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� Challenge Strategy: � Hold Times
� Fractional Cycle Approach
� No. of Lots Tested
� Product/Equipment Matrix Required
� Test Methods and Sampling Plan:
� Rinse Sampling
� Swab Sampling
� Visual Inspection
� Analytical Methods
� Assay Selection (Chemical/Micro)
� Assay Validated (Includes Swab Recovery)
� Acceptance Criteria
Defined in a Protocol and Includes:
Validation Strategy
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Overall surface evaluation: final rinse sampling & analysis
USP chemical- purified water methodology
pH 5.0 - 7.0
Conductivity < 3 µS/cm
Endotoxin < 0.25 EU/mL to < 10.00 EU/mL
TOC < 1.0 ppm+(based on system capability)
Bioburden* < 100 cfu/10mL
Product specific varies, typically non-detect
+ Over Negative Control
* Bioburden sampling is performed in systems that are not steamed or sterilized for
bioburden control
Typical Acceptance Criteria
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Back to Style-ogen® :Bulk Portable Tanks CIP
Next Use End of Use Cleaning
24 Hour
Dirty Hold Time 7 Day
Clean Hold Time
� Manufacturing Process:
� Product is Inactivated with Hypochlorite Prior to CIP (SOP)
� Only 100L Portable Tanks Cleaned at Bulk PTS
� One Tank Can Be Cleaned At A Time At Bulk PTS
� Each Tank Can Contain One of the Following Product Soils:
� Active Ingredient “Manny”
� Active Ingredient “Moe”
� Active Ingredient “Jack”
� 25% Sucrose
� Tank is Not SS/SIP After Cleaning
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Style-ogen® - BULK PTS CIP
� Cleaning Process:
� Automated CIP Cycle
� Cycle Steps Defined
� Cleaning SOP Available
� Cycle Development
Planned Concurrent to OQ & Engineering Lots
� Mfg. Equipment & Design:
� Bulk PTS & 100L Style-ogen® Tanks will be IQ/OQ’d
� All Product Contact Surfaces Constructed of Stainless Steel
� Good CIP Design Principles Followed
� Validation Sample Locations Readily Accessible &
Available (Rinse Sample Port)
Hold Time or Cycle
Step Description
Production
Cycle
Dirty Hold Time < 24 hrs.
HWFI Rinse 2 min.
2% Caustic Wash 15 min.
HWFI Rinse 2 min.
1% Acid Wash 5 min.
Final Rinse #1 2 min.
Final Rinse #2 2 min.
Final Rinse #3 2 min.
Final Rinse #4 2 min.
Final Rinse #5 2 min.
Clean Hold Time < 7 days
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Styleogen® - BULK PTS CIP
� Validation Strategy: Residual
Name
Time to Clean to
Acceptable Level
"Manny" 150
"Moe" 300
"Jack" 150
25% Sucrose 100
Lab Scale Cleanability Results
� How Many Validation Lots?
Hold Time or Cycle
Step Description
Production
Cycle
Validation
Cycle
Dirty Hold Time < 24 hrs. > 24 hrs.
HWFI Rinse 2 min. 2 min.
2% Caustic Wash 15 min. 10 min.
HWFI Rinse 2 min. 2 min.
1% Acid Wash 5 min. 5 min.
Final Rinse #1 2 min. 2 min.
Final Rinse #2 2 min. 2 min.
Final Rinse #3 2 min. 2 min.
Final Rinse #4 2 min. 2 min.
Final Rinse #5 2 min. N/A
Clean Hold Time < 7 days > 7 days
Cleaning Cycle Description � Use a Fractional Cycle Approach
� Caustic Wash Time Reduced 33%
� Final Rinse #5 Eliminated
� Dirty & Clean Hold Times
Challenged During Validation
� 3 Lots “Moe” (Hardest to Clean)
� 1 Lot Equivalency Each Others
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Style-ogen® - Bulk PTS CIP
� Execute Protocol – Document deviations
� Collect samples
� Analyze samples
� Check vs Acceptance Criteria – Pass / Fail / Investigation
� Write Report – Address deviations
� Review / Approve Report – Stakeholders � Include summary in license document
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Presentation Outline
� Validation Overview
� Cleaning Validation
� Process Validation
� Sterilization Validation
� Citation Examples
� Regulatory References
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Process Validation - Definition
[To establish] documented evidence which provides a
high degree of assurance that a specific process
will consistently produce a product meeting pre-determined specifications and quality attributes.
(FDA, May 1987)
2001-ms-3767
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� Demonstrate process control and consistency
� Comply with regulatory requirements for
licensure
� Provide assurance that release tests will be met;
the need for some release testing may be
eliminated.
Why Validate the Process ?
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Key Process Variables
Optimization/Process Understanding
Robustness Worst case challenges?
Process Validation at Full-scale
Process
Characterization
Process
Validation
Phase I/II Clinical
process
Lab-scale
process
Manufacturing
process
Lab Scale Validation
Process Validation requires a rational approach
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Characterization vs. Validation
� Characterization
– “Validation” studies at bench-scale using scaled-down
models, if possible.
– Well-documented in Lab notebooks and key technical
reports (no protocol)
– Learning, not “Validating”
� Validation
– Usually at Full-scale in actual process equipment (except
for viral clearance and resin/filter re-use)
– Conducted by Manufacturing under Protocol
– Testing what we already know, NOT EXPERIMENTING!
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Understand Your Process
� Ruggedness
– Multiple lots of raw materials
– Multiple lots of resins/filters
– Explore failure limits at laboratory/pilot scale
� Scaled-down process should reflect full
scale manufacturing performance as
closely as possible so that data generated
are relevant.
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Definitions
Critical Process Parameter (CPP):
An input variable that must be controlled within a specified
range to ensure success.
Critical Quality Attribute (CQA):
An output parameter from a unit operation that must be
within a specified range to demonstrate control,
consistency, and acceptable product quality.
CPP CQA
Ionic Strength Ion Exchange Yield
Column Load Chromatography Purity
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Example-Homogenization Step
Homogenization •Pressure
•Conc. •# of passes
•Temperature •Residence time
•Back-pressure
•Cell breakage
CPPs? CQA
Function in the manufacturing process:
Cell breakage - cell breakage must be ≥≥≥≥ 70% by Hematocrit assay.
• Process knowledge • Scientific rationale
•Tools are simply to provide a basis for discussion
and to facilitate the PV process.
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1. Select CPPs, CQAs
2. Process Validation Protocol
3. Execute
4. Assay
5. Report
6. File
Back to Style-ogen® : Process Validation
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Process Mapping: Step & Purpose
Fermentation
Thaw/Cell Breakage
Microfiltration/Chromatography 1
UF/Chromatography 2
Sterile Filtration
Dilution/Adjuvant Addition
Antigen Release
Antigen capture
Polishing purification
Sterilization
Dose/ adjuvant
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Detailed Step Description
Chromatography
Step Goal: Primary purification
Equipment constraints: •flow rate •pressure drop
•Cycle time •Column size •Flow distribution
Sampling plan: •Feed •Flow-through
•Product
Characterization •Size •Potency
•Lipid •Carbohydrates •Yield •Purity
Monitoring •Flow-pressure •UV
•Conductivity •HETP
Other parameters: Feed properties/composition, salt concentration, temperature, lot-to-lot feed/resin variability, feed concentration, load
Support Documents •Technical memos:
•Effect of load
•Cleaning/reuse •SOP’s •Batch summaries •Equipment FRS
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� Impact on product quality
– does the parameter have an impact on a CQA?
� Controllability
– how easy is the parameter to control?
� Recovery potential
– is there a redundant downstream step?
Use tools such as Criticality Index Analysis
Select CPPs, CQAs – Factors to Consider
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Example of a Criticality Index Analysis
Cell breakage
Enzyme
treatment
Microfiltration
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Process Validation Protocol
� CPPs, CQAs w/ acceptance criteria – Background / rationale for ranges
� How will they be sampled / monitored ?
� How many validation lots ?
� How will deviations be handled ?
Define Roles and Responsibilities
Manufacturing, Quality, Technology
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Process Validation Protocol
Step Goal CPPs CPP
Range
How
controll
ed
CQA Samples CQA
Range
Methods
Ferment
ation
High
cell
density
pH
Temp
7.0± 0.5 DCS Final
Glucose
Concn.
Broth –
final time
point
1 – 3
g/L
Analytical
methd
SOP XYZ
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Style-ogen® - Process Validation
� Complete 3 Validation Lots
� Obtain, Analyze data
� Address deviations �Transient deviations
�Equipment malfunctions
� Additional lots if needed
� Complete / approve report
� Include in license
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Presentation Outline
� Validation Overview
� Cleaning Validation
� Process Validation
� Sterilization Validation
� Citation Examples
� Regulatory References
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Sterilization Validation
� Sterilization Validation overview
� Validation of sterilization cycles (Protocol,
Acceptance criteria,=.)
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Steam Sterilization
� Cell death by protein denaturation
� Simple, reliable & economical
� Spores are more resistant than cells
� Spores ~100x more resistant to dry heat
than steam
� Typical cycle: >121ºC for 5-45 minutes
� Saturated steam is critical!
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Kinetics of Microbial Death
� Generally observed to be first-order
kinetics
� Non-logarithmic behavior is known
� Kinetic models
)( kt
o
oo
eNN
ttatNN
kNdt
dN
−=
==
=−
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Kinetics of Cell Death
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Kinetics of Cell Death
� Logarithmic decline most applicable to
vegetative cells
� Spores can show non-log rates
– Spore germination
– Sequential events for death
SSRR
S
RR
R
D
k
S
k
R
NkNkdt
dN
Nkdt
dN
NNN SR
−=
−=
→→
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Kinetics of Cell Death
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Temperature Effect
� Kinetic rate is a function of temperature
� Arrhenius model typically employed:
� Linear correlation between ln(k) and 1/T
)/( RTEeAk
∆−=
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Temperature Effect
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Characterization of Steam Sterilization Cycle Lethality
� Organism-related
– D-value (log reduction time)
– Z-value (deg. of temp. to reduce D by 1 log)
� Cycle-related
– F-value (integrated lethality delivered)
� Log reduction = F/D
– Typically, TR = 121.1º C, D = 1-3 min (spores)
– Target Fo = 36-72 minutes (full cycle)
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D-value
� A measure of the sensitivity of an
organism to a sterilization method
� Decimal reduction time - time at a given
temperature required to reduce a
population by 1 log
kDt
eN
N
eNN
kt
o
kt
o
303.2
10
1 )(
)(
==
==
=
−
−
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Z-value
� A measure of sensitivity of organism to different
temperatures
� Number of degrees needed to alter the D-value
by one log
� Allows for integration of the lethal effect of heat
as the temperature changes.
� Depends on sterilization method
– Steam: Z = 10º C
– Dry heat: Z = 21º C
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Z-value
� Spores, Z = 8 - 12ºC
� 10º C usually assumed
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F-value
� Integrated amount of lethality delivered
during a sterilization cycle
� For TR = 121º C and Z = 10º C, F = Fo
tFZTT R ∆∑
−=
/)(10
tFT
o∆∑
−=
10/)121(10
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F-value
� Extremely sensitive to temperature
– Fo = 1 min at 121º C for t = 1 min
– Fo = 2 min at 124º C for t = 1 min
– Fo = 8 min at 130º C for t = 1 min
� Log reduction = F/D
– D = 2 min, Fo = 16 min, Log reduction = 8
– For SAL = 10-6, initial population <102
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Typical SIP Cycle
� Come-up
– Purge air
– Add steam
– Wait to reach sterilization temperature
� Dwell
– Hold at T>121 C for fixed time or Fo
� Cool-down
– Turn steam off and cool system down
– Pressurize with air
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Sterilization Validation
� Fractional cycle approach
� Challenge with 106 G. stearothermophilus spores
� Run validation studies to obtain a 6 log reduction of
G. stearo. spores
� Production cycle will be based on a theoretical 12
log reduction
� Establish continuing validation schedule and change
control for validated cycle.
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Sterilization Validation (SIP)
� Place spore challenges throughout the system targeting
worst case locations (Geobacillus stearothermophilus)
� Run a fractional sterilization cycle (reduced temperature
and/or time)
� Evaluate the temperatures (Fo) at each location
� Evaluate saturated steam conditions
� Evaluate the kill/inactivation of the spores
� Perform 3 fractional cycle studies followed by 1 production
cycle study
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Sterilization Validation – Positioning of Thermocouples
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Validation Complete – What Next?
Cleaning Dev.
Sterilization Dev.
Practice Lots
IQ/OQ/Facility/Utility Qualification
Validation Lots
YEAR
APPROVAL
Phase III
APPROVAL
1H Y4 2H Y4 1H Y5 2H Y5
Cleaning Valdn.
Sterilization Valdn.
File Licens
e
Start Change
Control
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Implement Change Control
� Changes happen=.
� Need to
– Document changes
– Assess impact on
validation
– Revalidate as necessary
– File as necessary
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Presentation Outline
� Validation Overview
� Cleaning Validation
� Process Validation
� Sterilization Validation
� Citation Examples
� Regulatory References
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Recent FDA Observations - Cleaning
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Recent FDA Observations - Sterilization
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Recent FDA Observations - Process
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Regulatory References
� FDA guidance documents – CMC Guidance
� http://www.fda.gov/cber/gdlns/cmcvacc.pdf
– Sterilization Validation � http://www.fda.gov/cber/gdlns/sterval.pdf
– Process Validation � http://www.fda.gov/cder/guidance/pv.htm
– PAT approach � http://www.fda.gov/cder/guidance/6419fnl.htm
� FDA guidance documents – CMC Guidance
� http://www.fda.gov/cber/gdlns/cmcvacc.pdf
– Sterilization Validation � http://www.fda.gov/cber/gdlns/sterval.pdf
– Process Validation � http://www.fda.gov/cder/guidance/pv.htm
– PAT approach � http://www.fda.gov/cder/guidance/6419fnl.htm