Basis Validation of Biomanufacturing Processes

1,394 views
1,185 views

Published on

Basis Validation of Biomanufacturing Processes

Published in: Science, Business, Technology
0 Comments
1 Like
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total views
1,394
On SlideShare
0
From Embeds
0
Number of Embeds
14
Actions
Shares
0
Downloads
39
Comments
0
Likes
1
Embeds 0
No embeds

No notes for slide
  • The block flow is a simple representation of the production process illustrating “unit operations” with simple blocks. Major divisions in the production process, i.e. up-stream, downstream, viral and non-viral are separated with dashed lines.
    Each of these unit operations may contain numerous pieces of equipment, and proceedures that will require validation. In addition, the unit operation will require validation that the material coming out of that unit operation meets predetermined specifications.
  • Basis Validation of Biomanufacturing Processes

    1. 1. Validation Of Pharmabio-Validation Of Pharmabio- manufacture Processesmanufacture Processes Angel L. Salaman, PhDAngel L. Salaman, PhD angelsalaman@yahoo.comangelsalaman@yahoo.com
    2. 2. What is Validation?What is Validation? Validation – An Essential Part of GMPs!Validation – An Essential Part of GMPs! Validation is the scientific study of a systemValidation is the scientific study of a system  To prove that the facility/system/equipment/method isTo prove that the facility/system/equipment/method is consistently doing what it is supposed to do (i.e., that theconsistently doing what it is supposed to do (i.e., that the process is under control).process is under control). – We want to make decisions based on good science andWe want to make decisions based on good science and not hunches and assumptions!not hunches and assumptions!  To determine the process variables and acceptable limits forTo determine the process variables and acceptable limits for these variables, and to set-up appropriate in-processthese variables, and to set-up appropriate in-process controls.controls. – Is it ok if the wash from a chromatography column is pHIs it ok if the wash from a chromatography column is pH 6.8 vs. 7.0 ?6.8 vs. 7.0 ?
    3. 3. FDA definition ofFDA definition of validationvalidation ““Validation is a process ofValidation is a process of demonstratingdemonstrating, through, through documenteddocumented evidenceevidence, that a process, procedure,, that a process, procedure, method, piece of equipment, ormethod, piece of equipment, or facility willfacility will consistentlyconsistently produce aproduce a product or result that meetsproduct or result that meets predetermined specificationspredetermined specifications andand quality attributesquality attributes.”.”
    4. 4. Quality AttributesQuality Attributes?? IdentityIdentity – 21 CFR 211.84 (d) at least one test shall be conducted to verify the identity of each21 CFR 211.84 (d) at least one test shall be conducted to verify the identity of each component of a drug product.component of a drug product. – Chemical, biological, ImmunologicalChemical, biological, Immunological – Raw materials, In-process intermediates, final products.Raw materials, In-process intermediates, final products.  SafetySafety – 21 CFR 600.3 (p) safety as the relative freedom from harmful effect to persons affected,21 CFR 600.3 (p) safety as the relative freedom from harmful effect to persons affected, directly or indirectly, by a product when prudently administered, taking into considerationdirectly or indirectly, by a product when prudently administered, taking into consideration the character of the product in relationship to the condition of the recipient at the time.the character of the product in relationship to the condition of the recipient at the time.  Activity of active ingredientsActivity of active ingredients  Activity of the excipients or additivesActivity of the excipients or additives  Activity of process related impuritiesActivity of process related impurities  EfficacyEfficacy – Effectiveness of the product in achieving its medicinal purpose (therapeutic, prophylactic,Effectiveness of the product in achieving its medicinal purpose (therapeutic, prophylactic, diagnostic). Gathered at phase II and Phase III trials.diagnostic). Gathered at phase II and Phase III trials.  PotencyPotency – 21 CFR 600.3 (s) specific ability or capacity of the product, as indicated by its appropriate21 CFR 600.3 (s) specific ability or capacity of the product, as indicated by its appropriate laboratory tests or by adequately controlled clinical data obtained through the administrationlaboratory tests or by adequately controlled clinical data obtained through the administration of the product in the manner indicated to effect the given result.of the product in the manner indicated to effect the given result.  PurityPurity – 21 CFR 600.3 (r) relative freedom from extraneous matters in the finished product, whether21 CFR 600.3 (r) relative freedom from extraneous matters in the finished product, whether or not harmful to the recipient or deleterious to the product.or not harmful to the recipient or deleterious to the product.  Cleaning ProceduresCleaning Procedures  StabilityStability – 21 CFR 211.137 (a) to assure that a drug product meets applicable standards of identity,21 CFR 211.137 (a) to assure that a drug product meets applicable standards of identity, quality, and purity at the time of use; it shall bear an expiration date determined by stabilityquality, and purity at the time of use; it shall bear an expiration date determined by stability testing. Drugs may use accelerated time studies, biologics must use real time studies.testing. Drugs may use accelerated time studies, biologics must use real time studies.  ConsistencyConsistency – The ability of the product and/or process to reliably possess specified quality attributes on anThe ability of the product and/or process to reliably possess specified quality attributes on an ongoing basis. 3 consecutive batches of product meeting predetermined specifications isongoing basis. 3 consecutive batches of product meeting predetermined specifications is accepted as proof that a process is consistent. However, in NDA data from up to twentyaccepted as proof that a process is consistent. However, in NDA data from up to twenty batches may be submitted.batches may be submitted.
    5. 5. BiomanufacturingBiomanufacturing  Biomanufacturing is a complex process involvingBiomanufacturing is a complex process involving multiple unit operations many of which aremultiple unit operations many of which are criticalcritical to insuring patient safety and product efficacyto insuring patient safety and product efficacy
    6. 6. Inoculums Seed Fermentation Production Fermentation Harvest Ultrafiltration 1 Chrom. 1 1 Ultrafiltration 2 Chrom. 2 Viral Filtration Chrom. 3 Ultrafiltration 3 Final Formulation/ Sterile Filtration Sterile Fill UPSTREAM DOWN- STREAM VIRAL NON-VIRAL Block Flow Diagram of a typical Production Process
    7. 7. Historical Basis forHistorical Basis for ValidationValidation  AssumptionsAssumptions concerning virus inactivationconcerning virus inactivation resulted in ten deaths and 200 childrenresulted in ten deaths and 200 children becoming paralyzed, from a supposedlybecoming paralyzed, from a supposedly “inactivated” polio vaccine.“inactivated” polio vaccine.  AssumptionsAssumptions about sterilization causedabout sterilization caused severe infections among burn victims givensevere infections among burn victims given supposedly sterile solutions.supposedly sterile solutions.  Validation eliminates assumptions and reliesValidation eliminates assumptions and relies on experimental proof!on experimental proof!
    8. 8. Quality by DesignQuality by Design  A central concept in quality is thatA central concept in quality is that quality can not bequality can not be tested fortested for.. Quality must be designed and built into theQuality must be designed and built into the production process.production process. – Requires careful attention to raw materialRequires careful attention to raw material specificationsspecifications, in process material, in process material specificationsspecifications,, and final productand final product specificationsspecifications..
    9. 9. Validation andValidation and QualityQuality  Validating the performance of unit operations,Validating the performance of unit operations, analytical methods, and critical process pointsanalytical methods, and critical process points (sterilization, viral inactivation, cleaning(sterilization, viral inactivation, cleaning procedures) is essential in insuring that theprocedures) is essential in insuring that the process generates a quality product.process generates a quality product.
    10. 10. Validation in BiomanufacturingValidation in Biomanufacturing  Validation does not replace testing, but it doesValidation does not replace testing, but it does reduce the testing burden for raw materials,reduce the testing burden for raw materials, in-process materials, and final productin-process materials, and final product
    11. 11.  Validation itself is a process that evolves withValidation itself is a process that evolves with the product.the product. – Validation requirements for production ofValidation requirements for production of pre-clinical material less stringent then forpre-clinical material less stringent then for phase III clinical material.phase III clinical material. – Critical operations must be validatedCritical operations must be validated: For: For example: raw materials, analyticalexample: raw materials, analytical methods, viral clearance, sterilization,methods, viral clearance, sterilization, cleaning.cleaning. Validation in BiomanufacturingValidation in Biomanufacturing
    12. 12.  A fully validated process is “locked in”A fully validated process is “locked in”  Any change outside of the validated spaceAny change outside of the validated space invalidates processinvalidates process  Change must be evaluated for effect on patientChange must be evaluated for effect on patient safety and product efficacy-safety and product efficacy-Change control !Change control ! Validated Production Process Δ Validation in BiomanufacturingValidation in Biomanufacturing
    13. 13. Less More Difficulty in Changing and Binding Authority LAWS Regulation s Guidance Less More LAWS Regulation s Guidance Scientific Content & Detail about Implementation
    14. 14. Regulatory requirementsRegulatory requirements for validationfor validation 21 CFR 211 Subpart H- Holding and Distribution21 CFR 211 Subpart H- Holding and Distribution – 211.165 – Testing and release for211.165 – Testing and release for distributiondistribution  ““Requires that the accuracy, sensitivity,Requires that the accuracy, sensitivity, specificity, and reproducibility of testspecificity, and reproducibility of test methods employed by the firm shall bemethods employed by the firm shall be established and documented.established and documented. Such validationSuch validation and documentation may be accomplished inand documentation may be accomplished in accordance with 21 CFR 211.194 (a)(2)”accordance with 21 CFR 211.194 (a)(2)”
    15. 15.  Sec. 211.113 Control of microbiologicalSec. 211.113 Control of microbiological contamination.contamination.  (a) Appropriate written procedures, designed to(a) Appropriate written procedures, designed to prevent objectionable microorganisms in drugprevent objectionable microorganisms in drug products not required to be sterile, shall beproducts not required to be sterile, shall be established and followed.established and followed.  (b) Appropriate written procedures, designed to(b) Appropriate written procedures, designed to prevent microbiological contamination of drugprevent microbiological contamination of drug products purporting to be sterile, shall be establishedproducts purporting to be sterile, shall be established and followed.and followed. Such procedures shall include validationSuch procedures shall include validation of any sterilization process.of any sterilization process. Regulatory requirementsRegulatory requirements for validationfor validation
    16. 16. What does “validation of anyWhat does “validation of any sterilization process” mean ?sterilization process” mean ?  What parameters are critical to sterilization?What parameters are critical to sterilization? – Temperatures, pressures, time, pore sizeTemperatures, pressures, time, pore size (filtration), radiation dosage, chemical(filtration), radiation dosage, chemical concentration.concentration.  Must demonstrate that your autoclave reaches theMust demonstrate that your autoclave reaches the temperatures, pressures, and times necessary fortemperatures, pressures, and times necessary for sterilization.sterilization.  Must demonstrate that items representing real worldMust demonstrate that items representing real world samples achieve those conditions ( 20 ft of 1 ½ hose;samples achieve those conditions ( 20 ft of 1 ½ hose; a 20 L carboy; a 500 ml bottle).a 20 L carboy; a 500 ml bottle).  Must challenge with worse case scenario (may takeMust challenge with worse case scenario (may take place in pilot plant if scalability demonstrated).place in pilot plant if scalability demonstrated).
    17. 17. Regulatory guidance onRegulatory guidance on validationvalidation  Guideline on General Principals of Process ValidationGuideline on General Principals of Process Validation http://www.fda.gov/cder/guidance/pv.htmhttp://www.fda.gov/cder/guidance/pv.htm  Guidance for Industry: For the Submission DocumentationGuidance for Industry: For the Submission Documentation for Sterilization Process Validation in Applications for Humanfor Sterilization Process Validation in Applications for Human and Veterinary Drug Products. CDER CVM November 1994.and Veterinary Drug Products. CDER CVM November 1994. www.fda.gov/CDER/GUIDANCE/cmc2.pdfwww.fda.gov/CDER/GUIDANCE/cmc2.pdf  Working Party on Control of Medicines and InspectionsWorking Party on Control of Medicines and Inspections  Final Version of Annex 15Final Version of Annex 15 to the EU Guide to Goodto the EU Guide to Good Manufacturing PracticeManufacturing Practice  Title: Qualification and validationTitle: Qualification and validation  http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs- en/v4an15.pdfen/v4an15.pdf  ICH Q7a Section 12 on validationICH Q7a Section 12 on validation  http://www.fda.gov/cder/meeting/ICH_Q7A/index.htmhttp://www.fda.gov/cder/meeting/ICH_Q7A/index.htm  A WHO guide to good manufacturing practice (GMP)A WHO guide to good manufacturing practice (GMP) requirements. Part 2: Validationrequirements. Part 2: Validation  Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997.Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World Health Organization, Geneva.World Health Organization, Geneva.
    18. 18. Critical Operations inCritical Operations in BiomanufacturingBiomanufacturing  Some operations are more critical than others.Some operations are more critical than others. – Viral filtration, sterilization, cleaning, analyticalViral filtration, sterilization, cleaning, analytical methods.methods. – These operations will require greater validationThese operations will require greater validation efforts then less critical operations (mediaefforts then less critical operations (media blending).blending).
    19. 19. TestingTesting  Usually done by the Quality ControlUsually done by the Quality Control LaboratoryLaboratory – CFR requires thatCFR requires that quality unitquality unit be underbe under independent supervision and reportindependent supervision and report directly to senior managementdirectly to senior management
    20. 20. QualityQuality AssuranceAssurance  Reviews records from quality control and productionReviews records from quality control and production departmentsdepartments – Verifies that all specifications and productionVerifies that all specifications and production operations met / performedoperations met / performed – Investigations necessary for any deviationsInvestigations necessary for any deviations  Root causeRoot cause  Affect on qualityAffect on quality  Corrective action (CAPA)Corrective action (CAPA) – Approves final release of productApproves final release of product
    21. 21. Designing Quality intoDesigning Quality into the Productthe Product  Design of production process and specificationsDesign of production process and specifications all contribute to a quality product:all contribute to a quality product: – Absence of contaminationAbsence of contamination  Clean rooms, closed systems, use of BSCClean rooms, closed systems, use of BSC for critical operations.for critical operations. – PurityPurity  Separation process (chromatography)Separation process (chromatography) designed to remove potentialdesigned to remove potential contaminantscontaminants  Viral purification / inactivationViral purification / inactivation
    22. 22. ValidationValidation PlanPlan  Organizations must define anOrganizations must define an approach towards validationapproach towards validation – What is to be validatedWhat is to be validated – How is it to be validatedHow is it to be validated – Who is to validate itWho is to validate it – Who is to approve the validationWho is to approve the validation – When it must be revalidatedWhen it must be revalidated
    23. 23. ValidationValidation Examples of individualExamples of individual systems subject tosystems subject to validation:validation: HVAC systemsHVAC systems AutoclavesAutoclaves pH meterspH meters Depyrogenation OvensDepyrogenation Ovens LyopholyzersLyopholyzers CentrifugesCentrifuges Steam generatorsSteam generators Water systemsWater systems Compressed air systemsCompressed air systems Vacuum systemsVacuum systems
    24. 24. Validation PlanValidation Plan  Regulatory agencies (FDA, EMEA, WHO, etc) identifyRegulatory agencies (FDA, EMEA, WHO, etc) identify minimum components of validation.minimum components of validation.  ““Industry standards” (the c in cGMP) can increaseIndustry standards” (the c in cGMP) can increase validation requirements.validation requirements.  New & Novel processes / equipment require greaterNew & Novel processes / equipment require greater scrutiny then established processes / equipment.scrutiny then established processes / equipment.  Validation requirements increase as a product movesValidation requirements increase as a product moves through development (phase I, phase II, phase III).through development (phase I, phase II, phase III).
    25. 25. Validation PlansValidation Plans The Validation Master PlanThe Validation Master Plan – A high level document that outlines theA high level document that outlines the organizations philosophical approach to validationorganizations philosophical approach to validation and revalidation. The master validation planand revalidation. The master validation plan becomes a guideline by which individual validationbecomes a guideline by which individual validation protocol are developed and implemented.protocol are developed and implemented. – May contain a flow chart or other diagram of theMay contain a flow chart or other diagram of the validation processvalidation process
    26. 26. A prospectiveA prospective validation studyvalidation study IQ OQ Calibration PQ protocol approval PQ protocol execution Data Analysis Validation Report Approve Conclusions
    27. 27. DevelopmentalDevelopmental studiesstudies  Experiments designed to explore and define the limits ofExperiments designed to explore and define the limits of the system to be validatedthe system to be validated – Sterilization developmental studies may focus onSterilization developmental studies may focus on “worst case ” or hard to sterilize items“worst case ” or hard to sterilize items – Cleaning developmental studies may focus on “worstCleaning developmental studies may focus on “worst case” or hard to clean itemscase” or hard to clean items – Analytical methods may focus on defining the limitsAnalytical methods may focus on defining the limits of the procedure (range, recovery, etc)of the procedure (range, recovery, etc) – Developmental studies then used to developDevelopmental studies then used to develop validation protocols and refine SOP’svalidation protocols and refine SOP’s
    28. 28. Validation ProtocolValidation Protocol  Specific protocols (SOP’s) that provide detailedSpecific protocols (SOP’s) that provide detailed information on what is to be validated.information on what is to be validated.  Validation Protocols consist of:Validation Protocols consist of: – A description of the process, equipment, or methodA description of the process, equipment, or method to be validated.to be validated. – A description of the validation method.A description of the validation method. – A description of the sampling procedure including theA description of the sampling procedure including the kind and number of samples.kind and number of samples. – Acceptance criteria for test results.Acceptance criteria for test results. – Schedule or criteria for revalidation.Schedule or criteria for revalidation.
    29. 29. Example of a protocol for the IQ component of validating a pH meter As with all other SOP’s this document will contain an Objective, scope, and responsibility Section.
    30. 30. Validation ProtocolValidation Protocol  Validation Protocols may consist of multipleValidation Protocols may consist of multiple SOP’s each describing specific steps in theSOP’s each describing specific steps in the validation processvalidation process
    31. 31. Critical SystemsCritical Systems  How critical is the system being validated to finalHow critical is the system being validated to final product quality?product quality? – Media blending systems for cell growth vs. final fill &Media blending systems for cell growth vs. final fill & finish operationsfinish operations  Demonstrating that the device which fills, labels,Demonstrating that the device which fills, labels, and caps the final product will require moreand caps the final product will require more extensive validation then the blenders used toextensive validation then the blenders used to prepare media for bioreactors.prepare media for bioreactors.  Validation of complex devices may take years!Validation of complex devices may take years!
    32. 32. ValidationValidation  Proceeds in stages with new facilities /Proceeds in stages with new facilities / equipment.equipment.  Planning for validation should start withPlanning for validation should start with the design process.the design process.  Leaving validation to the last minute isLeaving validation to the last minute is asking for trouble.asking for trouble.
    33. 33. Stages of ValidationStages of Validation  Starts with Design & Receipt:Starts with Design & Receipt: – Does the equipment meet the needs (is theDoes the equipment meet the needs (is the autoclave big enough?)autoclave big enough?) – Do you have the manuals, spare parts, can youDo you have the manuals, spare parts, can you plug it in?plug it in? – Is it installed properly (drain lines, vents, etc)Is it installed properly (drain lines, vents, etc)  Does it work?Does it work? – Does the autoclave reach the necessary temp.Does the autoclave reach the necessary temp. and pressure?and pressure? – Can the autoclave sterilize your equipmentCan the autoclave sterilize your equipment (worse case situation)?(worse case situation)?  How does it work in the manufacturing process?How does it work in the manufacturing process? – Can it handle production quantities?Can it handle production quantities? – Will failure compromise product quality?Will failure compromise product quality?
    34. 34. IQ, OQ, PQ ?IQ, OQ, PQ ? Installation Qualification (IQ)Installation Qualification (IQ) A process used to document that the piece of equipmentA process used to document that the piece of equipment was supplied and installed properly and that appropriatewas supplied and installed properly and that appropriate utilities, i.e., electrical, steam, gas, etc. are available toutilities, i.e., electrical, steam, gas, etc. are available to operate the equipment according to the manufacturersoperate the equipment according to the manufacturers specifications.specifications. Operational Qualification (OQ)Operational Qualification (OQ) A process designed to supply the documented evidenceA process designed to supply the documented evidence that a piece of equipment operates as it is intendedthat a piece of equipment operates as it is intended through all anticipated operational ranges.through all anticipated operational ranges. Performance (Process) Qualification (PQ)Performance (Process) Qualification (PQ) Verifies that a process / piece of equipment performs as itVerifies that a process / piece of equipment performs as it is intended to in the manufacturing process and producesis intended to in the manufacturing process and produces product (in process or final) meeting predeterminedproduct (in process or final) meeting predetermined specifications.specifications.
    35. 35. Typical information in anTypical information in an IQ protocolIQ protocol  Name and description of equipment, including modelName and description of equipment, including model numbersnumbers  Identification, including model and serial numbersIdentification, including model and serial numbers  Location of the equipmentLocation of the equipment  Any utility requirements, i.e. electrical voltage, steamAny utility requirements, i.e. electrical voltage, steam or water pressure, etc.or water pressure, etc.  Any safety features of the equipment, includingAny safety features of the equipment, including alarms, interlocks, or relief valves.alarms, interlocks, or relief valves.  That all documentation, including manufacturersThat all documentation, including manufacturers contact information, spare parts inventory, operationalcontact information, spare parts inventory, operational manual, and installation drawings are available onmanual, and installation drawings are available on site.site.
    36. 36.  ObjectiveObjective  ResponsibilityResponsibility  Equipment required (Calibration verification &Equipment required (Calibration verification & Traceability)Traceability)  SOP(s) usedSOP(s) used  Equipment IdentificationEquipment Identification  Parameters measured (Specifications)Parameters measured (Specifications)  DocumentationDocumentation Typical information in anTypical information in an IQ protocolIQ protocol
    37. 37. ValidationValidation  Ideally validation takes place prior to actual production runs,Ideally validation takes place prior to actual production runs, however in some cases validation may take place as producthowever in some cases validation may take place as product is produced, or past production runs may be used to provideis produced, or past production runs may be used to provide validation data.validation data.  Prospective ValidationProspective Validation  Concurrent ValidationConcurrent Validation  Retrospective ValidationRetrospective Validation
    38. 38. RevalidationRevalidation  Is the initial validation of a piece of equipment the end?Is the initial validation of a piece of equipment the end? – No!No! – Periodic revalidation may be necessary depending onPeriodic revalidation may be necessary depending on the criticality of the equipmentthe criticality of the equipment – Changes need to be evaluated for their impact onChanges need to be evaluated for their impact on validationvalidation – Deviations from specifications may require revalidationDeviations from specifications may require revalidation – Revalidation spelled out in Master Validation PlanRevalidation spelled out in Master Validation Plan
    39. 39. Change ControlChange Control  Must assess impact of changes on FDA complianceMust assess impact of changes on FDA compliance and validation state.and validation state.  Change control is a formal process defined in companyChange control is a formal process defined in company SOP on how process/equipment changes areSOP on how process/equipment changes are evaluated.evaluated.  Any change that takes place outside the changeAny change that takes place outside the change control process can jeopardize product quality (patientcontrol process can jeopardize product quality (patient safety).safety).
    40. 40. Autoclave ValidationAutoclave Validation  IQ-IQ- – Design specifications meet users needsDesign specifications meet users needs – Proper installation, utilities, manuals,Proper installation, utilities, manuals, spare partsspare parts
    41. 41. The DQ, IQ, OQ process insures that this autoclave will meet the needs of the manufacturing group.
    42. 42. Sample Format for Installation Qualification of an autoclave. Courtesy of WHO. Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirements Part 2: Validation . World Health Organization, Geneva . www.who.int/vaccines-documents/DocsPDF/www9666.pdf Accessed on October 2nd, 2006.
    43. 43. Autoclave ValidationAutoclave Validation  OQOQ – Does it operate properlyDoes it operate properly  Does it reach the specified temperatureDoes it reach the specified temperature and pressureand pressure  Do timers workDo timers work  Does the operator interface panel workDoes the operator interface panel work  Are safety interlocks functionalAre safety interlocks functional
    44. 44. Sample Format for Operational Qualification of an autoclave. Courtesy of WHO. Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirements Part 2: Validation . World Health Organization, Geneva . www.who.int/vaccines-documents/DocsPDF/www9666.pdf A ccessed on October 2nd, 2006.
    45. 45. CalibrationCalibration Figure 1: Operational qualification of an autoclave requires the calibration of instruments against traceable standards. This figure shows the calibration of a validator (out of view) against the IRTD probe.
    46. 46. Monitoring TemperatureMonitoring Temperature Figure 2: A validator, used in the operational qualification of an autoclave. The validator is attached to individual thermocouples by wires coming from the rear of the instrument (arrow). Tha validator has been previously calibrated and the data gathered from the thermocouples will be logged on the laptop computer. The software on the computer is also subject to validation requirements.
    47. 47. The validator is attached to individual thermocouples (TC) by thin wires that pass through the wall of the autoclave through a specially designed port (arrow). This picture shows the back side of the autoclave. The validator is out of view at the lower left.
    48. 48. The inside of the autoclave showing the maze of wiring connecting the individual TC’s to the validator. The port through which the wires pass is visible in the middle left of the picture. The individual TC’s will be placed in various areas of the autoclave or equipment being autoclaved to generate a thermal map of the interior of the autoclave.
    49. 49. Output from the validator. The temperature at each connected thermocouple is displayed. Accumulated lethality (F 0 ) may also be displayed. Notice how some TC have failed and will not record a temperature. Accounting for TC failure is necessary to keep from having to repeat a study.
    50. 50. Proving that sufficient lethality (F0) is achieved within the nooks and crannies of biomanufacturing equipment requires the placement of TC’s in hard to reach areas, in this case deep within a piece of tubing. Special gaskets with openings for the TC are used to insert the TC within pieces of equipment.
    51. 51. Developmental studiesDevelopmental studies  Experiments designed to explore and define theExperiments designed to explore and define the limits of the system to be validatedlimits of the system to be validated – Sterilization developmental studies may focusSterilization developmental studies may focus on “worst case ” or hard to sterilize itemson “worst case ” or hard to sterilize items – Cleaning developmental studies may focus onCleaning developmental studies may focus on “worst case” or hard to clean items“worst case” or hard to clean items – Analytical methods may focus on defining theAnalytical methods may focus on defining the limits of the procedure (range, recovery, etc)limits of the procedure (range, recovery, etc)  Developmental studies then used to developDevelopmental studies then used to develop validation protocols and refine SOP’svalidation protocols and refine SOP’s
    52. 52. Developmental StudiesDevelopmental Studies Developmental studies are used to identify hard to autoclave items and to test if item preparation has an effect on ability to be sterilized
    53. 53. Developmental StudiesDevelopmental Studies Every little nook and cranny Needs to be assessed ! We have to prove that the inside of the pipe reaches sufficient temperature, for a long enough time to insure sterility!
    54. 54. Developmental StudiesDevelopmental Studies Does bagging make a difference?
    55. 55. Cleaning ValidationCleaning Validation Validating the cleaning cycle on a loaded dishwasher. Notice the various pipes and parts with narrow openings. Identification of hard to clean and easy to clean areas starts with developmental studies
    56. 56. How do we validate aHow do we validate a dishwasher ?dishwasher ?  How do we identify hard to clean & easy toHow do we identify hard to clean & easy to clean items?clean items?  How do we test to see if they are cleaned?How do we test to see if they are cleaned?
    57. 57. Going over the documentation
    58. 58. Cleaning ValidationCleaning Validation Sampling, documenting, and verifying is a labor intensive process Why is cleaning considered a critical process?
    59. 59. SamplingSampling Swabbing is commonly used to sample the surface of cleaned materials. The swab is then placed in a sample vial and sent to the Quality Control lab for analysis. Proper technique is essential in order to evaluate the effectiveness of cleaning techniques. Even so, swabbing results are typically corrected for known deficiencies in recovery.
    60. 60. CleaningCleaning ValidationValidation Analysis of rinse water for residual cleaning agents or process materials is an essential component of cleaning validation. Insuring that the sample is not contaminated requires vigilance and properly following the relevant SOP’s.
    61. 61. Analytical methods validationAnalytical methods validation  Considered a critical step in the manufacturing processConsidered a critical step in the manufacturing process – Requirements for validated analytical methodsRequirements for validated analytical methods explicitly written into the CFR’sexplicitly written into the CFR’s – 211.165 – Testing and release for distribution211.165 – Testing and release for distribution  ““Requires that the accuracy, sensitivity, specificity, andRequires that the accuracy, sensitivity, specificity, and reproducibility of test methods employed by the firmreproducibility of test methods employed by the firm shall be established and documented. Such validationshall be established and documented. Such validation and documentation may be accomplished in accordanceand documentation may be accomplished in accordance with 21 CFR 211.194 (a)(2)”with 21 CFR 211.194 (a)(2)”
    62. 62. Testing For IdentityTesting For Identity  Requires the development of validated analyticalRequires the development of validated analytical methods that can determine identity.methods that can determine identity.  Chemical Tests:Chemical Tests: – Is the molecule chemically what it is supposed toIs the molecule chemically what it is supposed to be?be?  Biological Activity Tests:Biological Activity Tests: – Does the molecules have theDoes the molecules have the biologic activitybiologic activity that itthat it is supposed to have?is supposed to have?  Immunogenic Tests:Immunogenic Tests: – Is the molecule immunogenic (allergic)?Is the molecule immunogenic (allergic)?
    63. 63. IdentityIdentity  21 CFR requires testing of raw materials:21 CFR requires testing of raw materials: – Raw materials quarantined until identity verifiedRaw materials quarantined until identity verified – Raw materials must meetRaw materials must meet predeterminedpredetermined specificationsspecifications – Vendors (and alternates) specified in BLAVendors (and alternates) specified in BLA (NDA)(NDA)
    64. 64. IdentityIdentity  21 CFR requires testing of in-process21 CFR requires testing of in-process materials:materials: – Product from bioreactor / fermentorProduct from bioreactor / fermentor – Product from purification stepsProduct from purification steps – Waste products from aboveWaste products from above Must meet specifications, if not - stopMust meet specifications, if not - stop thethe process to investigate take correctiveprocess to investigate take corrective actionaction
    65. 65. Regulatory guidance onRegulatory guidance on validationvalidation  Guideline on General Principals of Process ValidationGuideline on General Principals of Process Validation http://www.fda.gov/cder/guidance/pv.htmhttp://www.fda.gov/cder/guidance/pv.htm  Guidance for Industry: For the Submission DocumentationGuidance for Industry: For the Submission Documentation for Sterilization Process Validation in Applications for Humanfor Sterilization Process Validation in Applications for Human and Veterinary Drug Products. CDER CVM November 1994.and Veterinary Drug Products. CDER CVM November 1994. www.fda.gov/CDER/GUIDANCE/cmc2.pdfwww.fda.gov/CDER/GUIDANCE/cmc2.pdf  Working Party on Control of Medicines and InspectionsWorking Party on Control of Medicines and Inspections  Final Version of Annex 15Final Version of Annex 15 to the EU Guide to Goodto the EU Guide to Good Manufacturing PracticeManufacturing Practice  Title: Qualification and validationTitle: Qualification and validation  http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs- en/v4an15.pdfen/v4an15.pdf  ICH Q7a Section 12 on validationICH Q7a Section 12 on validation  http://www.fda.gov/cder/meeting/ICH_Q7A/index.htmhttp://www.fda.gov/cder/meeting/ICH_Q7A/index.htm  A WHO guide to good manufacturing practice (GMP)A WHO guide to good manufacturing practice (GMP) requirements. Part 2: Validationrequirements. Part 2: Validation  Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997.Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World Health Organization, Geneva.World Health Organization, Geneva.
    66. 66. Validation ProtocolValidation Protocol  Specific protocol based on developmental studiesSpecific protocol based on developmental studies  Protocol is written, reviewed and approvedProtocol is written, reviewed and approved  Protocol is executedProtocol is executed  Report written and approvedReport written and approved  System is validatedSystem is validated
    67. 67. Analytical Methods ValidationAnalytical Methods Validation  Being a critical component of productionBeing a critical component of production process analytical methods must beprocess analytical methods must be validatedvalidated – Raw material testingRaw material testing – In process materialsIn process materials – Final product specificationsFinal product specifications
    68. 68. What must beWhat must be demonstrateddemonstrated  Selectivity (specificity)Selectivity (specificity)  AccuracyAccuracy  PrecisionPrecision  Linear RangeLinear Range  Limit of detection (LOD)Limit of detection (LOD)  Limit of quantification (LOQ orLimit of quantification (LOQ or LLOQ)LLOQ)  RobustnessRobustness
    69. 69. ValidatedValidated methodsmethods  USP NF (United States Pharmacopeia NationalUSP NF (United States Pharmacopeia National Formulary) contains “validated analytical methods”Formulary) contains “validated analytical methods”  Use of a USP method does not eliminate theUse of a USP method does not eliminate the organizations obligation to demonstrate that theorganizations obligation to demonstrate that the method performs adequetlymethod performs adequetly
    70. 70. SelectivitySelectivity (specificity)(specificity)  Does the analytical method detect the component it is supposedDoes the analytical method detect the component it is supposed to detect?to detect? – Cross reactivity in antibody based methodsCross reactivity in antibody based methods  Demonstrate specificity by conducting analytical method onDemonstrate specificity by conducting analytical method on materials that may mimic analyte of interestmaterials that may mimic analyte of interest – Looking for “false positives”Looking for “false positives”
    71. 71. AccurAccur acyacy  Ability of analytical method to accurately determineAbility of analytical method to accurately determine the presence and amount of the analyte of interestthe presence and amount of the analyte of interest – Typically done by analyzing a traceable standardTypically done by analyzing a traceable standard
    72. 72. RecovRecov eryery  Can we recover all of the analyte from a complex matrixCan we recover all of the analyte from a complex matrix  May reflect sample preparation problemsMay reflect sample preparation problems  Typical recovery studies done using “spiked” samplesTypical recovery studies done using “spiked” samples  Final results may be corrected by the % recoveryFinal results may be corrected by the % recovery – Swab samples typically corrected to reflect recoverySwab samples typically corrected to reflect recovery
    73. 73. LinearLinear RangeRange  Must define the linear range of aMust define the linear range of a methodmethod – Assay may have multiple linearAssay may have multiple linear rangesranges
    74. 74. PrecisioPrecisio nn  How much variability does the assay exhibit when analyzingHow much variability does the assay exhibit when analyzing the same samplethe same sample – Typically demonstrated by analyzing multiple aliquots of aTypically demonstrated by analyzing multiple aliquots of a homogenous samplehomogenous sample – Acceptance criteria will depend on the assay and theAcceptance criteria will depend on the assay and the material being assayed (2-20% RSD)material being assayed (2-20% RSD) – Typically expressed as % RSD (relative standard deviation)Typically expressed as % RSD (relative standard deviation)
    75. 75. Limit of DetectionLimit of Detection (LOD)(LOD)  Lowest level at which method can detectLowest level at which method can detect analyteanalyte – Results reported as less than LODResults reported as less than LOD – Based on signal to noise specificationBased on signal to noise specification (10:1, 20:1)(10:1, 20:1)
    76. 76. RobustnessRobustness studiesstudies  How sensitive is the method to minor variations in methodHow sensitive is the method to minor variations in method – Pipetting variationPipetting variation – Temperature fluctuationTemperature fluctuation – Reagent stabilityReagent stability – Etc.Etc. Detailed robustness studies will be reflected in final SOP’sDetailed robustness studies will be reflected in final SOP’s
    77. 77. Limit ofLimit of Quantification (LOQ)Quantification (LOQ)  Lowest level at which method can accurately quantify analyteLowest level at which method can accurately quantify analyte – Based on signal-to-noise ratio specification (10:1, 20:1)Based on signal-to-noise ratio specification (10:1, 20:1) and precision specificationand precision specification  Precision and LOQ relatedPrecision and LOQ related – Lower LOQ will typically result in lower precisionLower LOQ will typically result in lower precision
    78. 78. BradfordBradford AssayAssay for totalfor total proteinprotein Well known colorimetric assay that relies on the binding ofWell known colorimetric assay that relies on the binding of Commassie G-250 dye to the proteins in an acidic solutionCommassie G-250 dye to the proteins in an acidic solution – Dye binding proportional to number of positive charges inDye binding proportional to number of positive charges in proteinprotein – Proteins >3000 dal not detectedProteins >3000 dal not detected  Simple, quick, wide range, few interfering agentsSimple, quick, wide range, few interfering agents
    79. 79. BradfordBradford AssayAssay
    80. 80. DisadvantaDisadvanta gesges  Incompatability with surfactantsIncompatability with surfactants  Staining of glass and quartz cuvettesStaining of glass and quartz cuvettes – Use disposable polystyrene cuvettesUse disposable polystyrene cuvettes – Or wash with strong detergents and methanolOr wash with strong detergents and methanol
    81. 81. Validating theValidating the BradfordBradford  Selectivity (specificity)Selectivity (specificity)  AccuracyAccuracy  RecoveryRecovery  PrecisionPrecision  Linear RangeLinear Range  Limit of detection (LOD)Limit of detection (LOD)  Limit of quantification (LOQ or LLOQ)Limit of quantification (LOQ or LLOQ)  RobustnessRobustness
    82. 82. Some QuestionsSome Questions  A valve used to transfer material from a holding tank to theA valve used to transfer material from a holding tank to the purification suite jam’s closed. You have a spare valve that ispurification suite jam’s closed. You have a spare valve that is an identical model. Can you change this valve with the sparean identical model. Can you change this valve with the spare and continue operations? What if the valve is from a differentand continue operations? What if the valve is from a different manufacturer?manufacturer?  You notice that your autoclave loading plan leaves room forYou notice that your autoclave loading plan leaves room for additional material. Realizing that increasing that amount ofadditional material. Realizing that increasing that amount of material in the autoclave will shorten the turn around timematerial in the autoclave will shorten the turn around time for the production line you contemplate increasing thefor the production line you contemplate increasing the amount of material loaded into the autoclave then specifiedamount of material loaded into the autoclave then specified by the loading plan. What should you do? What will beby the loading plan. What should you do? What will be required to implement this change?required to implement this change?  An SOP for calibration of a pH meter calls for a two pointAn SOP for calibration of a pH meter calls for a two point calibration at pH 4 and pH 7. You notice that a single pointcalibration at pH 4 and pH 7. You notice that a single point calibration at pH 7 produces the same result from pHcalibration at pH 7 produces the same result from pH measurements of your buffer solutions and allows you tomeasurements of your buffer solutions and allows you to take a longer break. Is it Ok to do the one point calibrationtake a longer break. Is it Ok to do the one point calibration when the SOP calls for a two point calibration? How wouldwhen the SOP calls for a two point calibration? How would you go about changing the SOP to allow for a one pointyou go about changing the SOP to allow for a one point calibration?calibration?
    83. 83.  What documents would provide information concerning theWhat documents would provide information concerning the make and model of a particular valve used to regulate themake and model of a particular valve used to regulate the transfer of material from a holding tank to the purificationtransfer of material from a holding tank to the purification suite?suite?  Your supervisor is concerned that the fermentation vesselYour supervisor is concerned that the fermentation vessel is not providing sufficient aeration of the culture to getis not providing sufficient aeration of the culture to get optimal growth and suggests installing a different kind ofoptimal growth and suggests installing a different kind of baffle in the vessel. How would you demonstrate that thisbaffle in the vessel. How would you demonstrate that this change has no effect on product quality?change has no effect on product quality?
    84. 84. ReferencesReferences  Pharmaceutical Manufacturers Association’s (Pharmaceutical Research and Manufacturers ofPharmaceutical Manufacturers Association’s (Pharmaceutical Research and Manufacturers of America) Validation Advisory Committee “Process Validation Concepts for Drug Products”America) Validation Advisory Committee “Process Validation Concepts for Drug Products” Pharmaceutical Technology, September 1985 p 82.Pharmaceutical Technology, September 1985 p 82.  Bismuth, G. Cleaning Validation: A Practical Approach. CRC Press, 2000. ISBN 1574911082.Bismuth, G. Cleaning Validation: A Practical Approach. CRC Press, 2000. ISBN 1574911082.  Pharmaceutical Process Validation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, MarcelPharmaceutical Process Validation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, Marcel Decker, 2003. ISBN 082470838-5Decker, 2003. ISBN 082470838-5  Validation of Pharmaceutical Processes: Sterile Products. 1998. 2nd Edition. Edited by Frederick J.Validation of Pharmaceutical Processes: Sterile Products. 1998. 2nd Edition. Edited by Frederick J. Carlton and James Agalloco. Marcel Decker, 1998. ISBN 0824793846.Carlton and James Agalloco. Marcel Decker, 1998. ISBN 0824793846.  Validation Standard Operating Procedures: A step by Step Guide for Achieving Compliance in theValidation Standard Operating Procedures: A step by Step Guide for Achieving Compliance in the Pharmaceutical, Medical Device, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002.Pharmaceutical, Medical Device, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002. ISBN 1574443313.ISBN 1574443313.  Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control FromGood Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control From Manufacturer to Consumer, Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.Manufacturer to Consumer, Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.  Voss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBNVoss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBN 0939459507.0939459507.  LeBlanc, D.A. 2000. Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRCLeBlanc, D.A. 2000. Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRC Press. ISBN 1574911163.Press. ISBN 1574911163.  Cloud, P. 1998. Pharmaceutical Equipment Validation: The Ultimate Qualification Guidebook. CRCCloud, P. 1998. Pharmaceutical Equipment Validation: The Ultimate Qualification Guidebook. CRC Press. ISBN 1574910795.Press. ISBN 1574910795.  Juran, Quality Control Handbook, 4th Edition., McGraw-Hill, 1988.Juran, Quality Control Handbook, 4th Edition., McGraw-Hill, 1988.  DeSain C, Sutton C. (1995). Process development that supports process validation.DeSain C, Sutton C. (1995). Process development that supports process validation. Pharmaceutical Technology 19 (Oct.): 130-136, 1995.Pharmaceutical Technology 19 (Oct.): 130-136, 1995.  Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess theGarcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a powder mixture.uniformity of a powder mixture. Drug Development and Industrial PharmacyDrug Development and Industrial Pharmacy 27(4): 297-307,27(4): 297-307, 2001.2001.  Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practiceChaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirements Part 2: Validation . World Health Organization, Geneva.(GMP) requirements Part 2: Validation . World Health Organization, Geneva. www.who.int/vaccines-documents/DocsPDF/www9666.pdf Accessed on October 2nd, 2006.www.who.int/vaccines-documents/DocsPDF/www9666.pdf Accessed on October 2nd, 2006.  Brown, F. 1993. Review of accidents caused by incomplete inactivation of viruses.Brown, F. 1993. Review of accidents caused by incomplete inactivation of viruses. Dev. Biol.Dev. Biol. Stand.Stand. 81:81: 103-7103-7  Nathanson, N. and Langmuir, A.D. 1995Nathanson, N. and Langmuir, A.D. 1995.. The Cutter incident. Poliomyelitis followingThe Cutter incident. Poliomyelitis following formaldehyde-inactivated poliovirus vaccination in the United States during the Spring of 1955. II.formaldehyde-inactivated poliovirus vaccination in the United States during the Spring of 1955. II. Relationship of poliomyelitis to Cutter vaccine. 1963.Relationship of poliomyelitis to Cutter vaccine. 1963. Am. J. Epidemiol.Am. J. Epidemiol. 142:109-40142:109-40..

    ×