Validation lecture

					Validation in Biotech Facilities:
     What ? Why ? How?




      Dr. PK Yegneswaran
    Presentation Outline

   Validation Overview
   Cleaning Validation
   Process Validation
   Sterilization Validation
   Citation Examples
   Regulatory References




    2                          2
           Typical Project Schedule
                                              YEAR
               1       2                  3                4                    5                6

                                               Phase III

                                                                                      APPROVAL
           Scope

                   Design

                            Procurement

                               Construction

                                                     IQ/OQ

                                                               Startup / Validation
100%                                                                                  APPROVAL
 % Spent




           3                                                                                         3
Typical Post OQ Schedule
                                                  YEAR


1H Y4                        2H Y4                               1H Y5              2H Y5

                                                       Phase III


    IQ/OQ/Facility/Utility Qualification                                                    APPROVAL

                                 Sterilization
                                     Dev.

                                           Sterilization
                                              Valdn.


                                                 Practice Lots


                                              Cleaning Dev.


                                                                                    File
                                                             Cleaning Valdn.
                                                                                   Licens
                                                                                      e
                                                                 Validation Lots
                                                                                            APPROVAL

4                                                                                                      4
 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)




                                                  5
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 !




6                                                     6
Why Validate?


                              Consistent yield &
                               quality
                              Rapid decisions when
                               mishaps occur
                              Fewer discards
                              Less time hosting
 … to ensure that the          government agencies,
 output is consistent;         more time
first time, every time!!       manufacturing
 7                                                7
What do we Validate ?


   Process
   Cleaning
   Sterilization
   Filters
   Containers
   Assays




    8                   8
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


    9                                                                        9
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
10                                                                     10
    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




    11                                                                   11
Typical Post OQ Schedule
                                                   YEAR


1H Y4                         2H Y4                               1H Y5              2H Y5

                                                        Phase III


     IQ/OQ/Facility/Utility Qualification                                                    APPROVAL

                                  Sterilization
                                      Dev.

                                            Sterilization
                                               Valdn.


                                                  Practice Lots


                                               Cleaning Dev.


                                                                                     File
                                                              Cleaning Valdn.
                                                                                    Licens
                                                                                       e
                                                                  Validation Lots
                                                                                             APPROVAL

12                                                                                                      12
    Presentation Outline

   Validation Overview
   Cleaning Validation
   Process Validation
   Sterilization Validation
   Citation Examples
   Regulatory References




13                             13
    Cleaning Validation

   Cleaning Validation overview
   Cycle development for Style-ogen®
    equipment
   Validation of cleaning cycles




14                                      14
What is Cleaning Validation ?

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.
15                                                15
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 product….”




  16                                                         16
  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.”


   17                                                     17
One-Third Of Recent Drug GMP Warning Letters
Cite Cleaning Practices (2002 survey)



Year        # of Warning   # of Warning      % Related to
            Letters        Letters related   Cleaning
                           to Cleaning

1999        65             23                35%


2000        71             20                41%


2001        71             20                28%




18                                                          18
Cleaning Validation
Where Do I Start?




     Manufacturing       Cleaning          Mfg. Equipment
       Process           Process              & Design




                     Validation Strategy
19                                                          19
 Manufacturing Process

Considerations:
  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 ?
 20                                                                              20
 Cleaning Process

Considerations:
 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?


  21                                                         21
  Mfg. Equipment Design

Considerations:
  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
 22                                                   22
  Validation Strategy

Defined in a Protocol and Includes:
  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)
 23
  Acceptance Criteria                                23
Typical Acceptance Criteria


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

    24                                                                           24
  Back to Style-ogen® :Bulk Portable Tanks CIP

 Manufacturing Process:
     End of Use                     Cleaning                     Next Use
                  24 Hour                      7 Day
                  Dirty Hold Time              Clean Hold Time


   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
    25                                                                      25
 Style-ogen® - BULK PTS CIP

 Cleaning Process:                   Hold Time or Cycle
                                      Step Description
                                                           Production
                                                           Cycle
    Automated CIP Cycle              Dirty Hold Time
                                      HWFI Rinse
                                                           < 24 hrs.
                                                           2 min.
    Cycle Steps Defined              2% Caustic Wash      15 min.
                                      HWFI Rinse           2 min.
    Cleaning SOP Available           1% Acid Wash         5 min.
                                      Final Rinse #1       2 min.
    Cycle Development                Final Rinse #2       2 min.
                                      Final Rinse #3       2 min.
     Planned Concurrent to            Final Rinse #4       2 min.
     OQ & Engineering Lots            Final Rinse #5       2 min.
                                      Clean Hold Time      < 7 days
 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)
 26                                                                     26
   Styleogen® - BULK PTS CIP

 Validation Strategy:                   Lab Scale Cleanability Results
                                             Residual     Time to Clean to
   How Many Validation Lots?                 Name        Acceptable Level
                                             "Manny"            150
      3 Lots “Moe” (Hardest to Clean)        "Moe"             300
                                              "Jack"            150
      1 Lot Equivalency Each Others       25% Sucrose          100


   Use a Fractional Cycle Approach        Cleaning Cycle Description
                                         Hold Time or Cycle Production Validation
      Caustic Wash Time Reduced 33%      Step Description    Cycle      Cycle
                                           Dirty Hold Time   < 24 hrs.  > 24 hrs.
      Final Rinse #5 Eliminated             HWFI Rinse       2 min.     2 min.
                                          2% Caustic Wash     15 min.   10 min.
                                             HWFI Rinse       2 min.     2 min.
   Dirty & Clean Hold Times               1% Acid Wash       5 min.     5 min.
                                            Final Rinse #1    2 min.     2 min.
   Challenged During Validation             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
    27                                     Clean Hold Time   < 7 days   > 7 days
                                                                           27
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



28                                           28
    Presentation Outline

   Validation Overview
   Cleaning Validation
   Process Validation
   Sterilization Validation
   Citation Examples
   Regulatory References




29                             29
                                            2001-ms-3767
 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)




                                                  30
      Why Validate the Process ?


   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.




31                                                  31
  Process Validation
  requires a rational approach

                       Key Process
                        Variables
                                                Lab-scale
                                                 process
                   Optimization/Process
Process
                      Understanding
Characterization
                                            Phase I/II Clinical
                                                 process
                       Robustness
                   Worst case challenges?



                   Lab Scale Validation
                                             Manufacturing
 Process                                        process
 Validation
     32            Process Validation at
                                                            32
                        Full-scale
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!



33                                                              33
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.


34                                                          34
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



35                                                               35
 Example-Homogenization Step

Function in the manufacturing process:
Cell breakage - cell breakage must be  70% by Hematocrit assay.

            CPPs?                                    CQA
         •Pressure
                           Homogenization           •Cell breakage
         •Conc.
         •# of passes
         •Temperature
         •Residence time
         •Back-pressure
                           • Process knowledge
                           • Scientific rationale
                           •Tools are simply to provide a basis
                           for discussion
   36                      and to facilitate the PV process.         36
Back to Style-ogen® : Process Validation

   1. Select   CPPs, CQAs

   2. Process Validation Protocol

   3. Execute

   4. Assay

   5. Report

   6. File

    37                              37
Process Mapping: Step & Purpose

           Fermentation


        Thaw/Cell Breakage           Antigen Release


  Microfiltration/Chromatography 1   Antigen capture


      UF/Chromatography 2            Polishing purification


          Sterile Filtration         Sterilization


     Dilution/Adjuvant Addition      Dose/ adjuvant
                                                              38
 Detailed Step Description

Step Goal: Primary purification


                               Chromatography

 Other parameters:
 Feed properties/composition, salt concentration, temperature, lot-to-lot feed/resin variability,
 feed concentration, load

Equipment constraints:   Sampling plan:   Characterization   Monitoring     Support Documents
•flow rate               •Feed            •Size              •Flow-pressure •Technical memos:
•pressure drop           •Flow-through    •Potency           •UV                 •Effect of load
•Cycle time              •Product         •Lipid             •Conductivity       •Cleaning/reuse
•Column size                              •Carbohydrates     •HETP          •SOP’s
•Flow distribution                        •Yield                            •Batch summaries
                                          •Purity                           •Equipment FRS
       39                                                                                39
Select CPPs, CQAs – Factors to Consider

     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


      40                                               40
  Example of a Criticality Index Analysis


Cell breakage


  Enzyme
  treatment



Microfiltration



    41                               41
    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



42                                           42
       Process Validation Protocol


Step        Goal   CPPs CPP          How        CQA      Samples     CQA     Methods
                        Range        controll                        Range
                                     ed

Ferment High       pH     7.0± 0.5   DCS        Final   Broth –      1–3     Analytical
ation   cell       Temp                         Glucose final time   g/L     methd
        density                                 Concn. point                 SOP XYZ




       43                                                                         43
 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




 44                                 44
    Presentation Outline

   Validation Overview
   Cleaning Validation
   Process Validation
   Sterilization Validation
   Citation Examples
   Regulatory References




45                             45
    Sterilization Validation

   Sterilization Validation overview
   Validation of sterilization cycles (Protocol,
    Acceptance criteria,….)




46                                              46
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!


47                                            47
Kinetics of Microbial Death



   Generally observed to be first-order
    kinetics
   Non-logarithmic behavior is known
   Kinetic models      dN
                               kN
                         dt
                       N  N o at t  t o
                       N  N o e ( kt )

48                                          48
Kinetics of Cell Death




49                       49
    Kinetics of Cell Death



   Logarithmic decline most applicable to
    vegetative cells
   Spores can show non-log rates
    –   Spore germination
    –   Sequential events for death
     kR       kS
             
N R   N S   N D
dN R
      kR N R
 dt
dN S
50  kR N R  k S N S
 dt                                          50
Kinetics of Cell Death




51                       51
    Temperature Effect



   Kinetic rate is a function of temperature
   Arrhenius model typically employed:
                   (  ΔE / RT )
          k  Ae
   Linear correlation between ln(k) and 1/T



52                                              52
     Temperature Effect




53                        53
    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)


54                                                        54
     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
             N  N o e ( kt )
             N    1
                    e ( kt )
             N o 10
                        2.303
             t D 
                          k
55                                             55
         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




56                                                        56
     Z-value


   Spores, Z = 8 - 12ºC
   10º C usually assumed




    57                      57
    F-value

   Integrated amount of lethality delivered
    during a sterilization cycle

      F  10   (T TR ) / Z
                               Δt
   For TR = 121º C and Z = 10º C, F = Fo

      Fo  10   (T 121 /10
                        )
                                Δt



58                                             58
     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




59                                                   59
    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

60                                                 60
    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.




61                                                      61
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
62                                                              62
Sterilization Validation – Positioning of
Thermocouples




63                                          63
Validation Complete – What Next?
                                                   YEAR


1H Y4                         2H Y4                               1H Y5              2H Y5

                                                        Phase III


     IQ/OQ/Facility/Utility Qualification                                                     APPROVAL

                                  Sterilization
                                      Dev.

                                            Sterilization
                                                                                     Start
                                               Valdn.
                                                                                    Change
                                                                                    Control
                                                  Practice Lots


                                               Cleaning Dev.


                                                                                     File
                                                              Cleaning Valdn.
                                                                                    Licens
                                                                                       e
                                                                  Validation Lots
                                                                                              APPROVAL

64                                                                                                       64
Implement Change Control

    Changes happen….
    Need to
     –   Document changes
     –   Assess impact on
         validation
     –   Revalidate as necessary
     –   File as necessary




65                                 65
    Presentation Outline

   Validation Overview
   Cleaning Validation
   Process Validation
   Sterilization Validation
   Citation Examples
   Regulatory References




66                             66
Recent FDA Observations - Cleaning




67                                   67
Recent FDA Observations -
Sterilization




68                          68
Recent FDA Observations - Process




69                                  69
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
             http://www.fda.gov/cder/guidance/pv.htm
    –   PAT approach
        PAT approach
            http://www.fda.gov/cder/guidance/6419fnl.htm




70                                                          70