Basis Validation of Biomanufacturing Processes

Validation Of Pharmabio-Validation Of Pharmabio-manufacture Processesmanufacture Processes

Angel L. Salaman, PhDAngel L. Salaman, [email protected]@yahoo.com

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 is consistently To prove that the facility/system/equipment/method is consistently

doing what it is supposed to do (i.e., that the process is under doing what it is supposed to do (i.e., that the process is under control).control).

– We want to make decisions based on good science and not We want to make decisions based on good science and not hunches and assumptions!hunches and assumptions!

To determine the process variables and acceptable limits for these To determine the process variables and acceptable limits for these variables, and to set-up appropriate in-process controls.variables, and to set-up appropriate in-process controls.

– Is it ok if the wash from a chromatography column is pH 6.8 vs. Is it ok if the wash from a chromatography column is pH 6.8 vs. 7.0 ?7.0 ?

FDA definition of FDA definition of validationvalidation

““Validation is a process of Validation is a process of demonstratingdemonstrating, , through through documented evidencedocumented evidence, that a , that a process, procedure, method, piece of process, procedure, method, piece of equipment, or facility will equipment, or facility will consistentlyconsistently produce a product or result that meets produce a product or result that meets predetermined specificationspredetermined specifications and and quality quality attributesattributes.”.”

Quality AttributesQuality Attributes? ? IdentityIdentity

– 21 CFR 211.84 (d) at least one test shall be conducted to verify the identity of each component of 21 CFR 211.84 (d) at least one test shall be conducted to verify the identity of each component of a drug product. 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, directly or 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 consideration the character of 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 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 appropriate 21 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 administration of laboratory tests or by adequately controlled clinical data obtained through the administration of the product in the manner indicated to effect the given result. 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, whether or not 21 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.harmful to the recipient or deleterious to the product. Cleaning Procedures Cleaning Procedures

StabilityStability– 21 CFR 211.137 (a) to assure that a drug product meets applicable standards of identity, quality, 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 stability testing. Drugs 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. 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 an The 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 is accepted ongoing 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 twenty batches may be as proof that a process is consistent. However, in NDA data from up to twenty batches may be submitted. submitted.

BiomanufacturingBiomanufacturing Biomanufacturing is a complex process involving Biomanufacturing is a complex process involving

multiple unit operations many of which are multiple unit operations many of which are criticalcritical to insuring patient safety and product efficacyto insuring patient safety and product efficacy

Inoculums Seed Fermentation

Production Fermentation Harvest

Ultrafiltration1

Chrom. 11

Ultrafiltration2

Chrom. 2

Viral Filtration

Chrom.3

Ultrafiltration3

Final Formulation/Sterile Filtration

Sterile Fill

UPSTREAM

DOWN-STREAM

VIRAL

NON-VIRAL

Block Flow Diagram of a typical Production Process

Historical Basis for Historical Basis for Validation Validation

Assumptions Assumptions concerning virus inactivation resulted concerning virus inactivation resulted in ten deaths and 200 children becoming paralyzed, in ten deaths and 200 children becoming paralyzed, from a supposedly “inactivated” polio vaccine. from a supposedly “inactivated” polio vaccine.

AssumptionsAssumptions about sterilization caused severe about sterilization caused severe infections among burn victims given supposedly infections among burn victims given supposedly sterile solutions. sterile solutions.

Validation eliminates assumptions and relies on Validation eliminates assumptions and relies on experimental proof!experimental proof!

Quality by DesignQuality by Design

A central concept in quality is that A central concept in quality is that quality can not be tested forquality can not be tested for. . Quality must be designed and built into the production process. Quality must be designed and built into the production process. – Requires careful attention to raw material Requires careful attention to raw material specificationsspecifications, in , in

process material process material specificationsspecifications, and final product , and final product specificationsspecifications. .

Validation and Validation and QualityQuality

Validating the performance of unit operations, analytical Validating the performance of unit operations, analytical methods, and critical process points (sterilization, viral methods, and critical process points (sterilization, viral inactivation, cleaning procedures) is essential in insuring inactivation, cleaning procedures) is essential in insuring that the process generates a quality product.that the process generates a quality product.

Validation in BiomanufacturingValidation in Biomanufacturing

Validation does not replace testing, but it does Validation does not replace testing, but it does reduce the testing burden for raw materials, in-reduce the testing burden for raw materials, in-process materials, and final productprocess materials, and final product

Validation itself is a process that evolves with Validation itself is a process that evolves with the product.the product.– Validation requirements for production of Validation requirements for production of

pre-clinical material less stringent then for pre-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, analytical methods, example: raw materials, analytical methods, viral clearance, sterilization, cleaning. viral clearance, sterilization, cleaning.


A fully validated process is “locked in” A fully validated process is “locked in” Any change outside of the validated space Any change outside of the validated space

invalidates process invalidates process Change must be evaluated for effect on patient Change must be evaluated for effect on patient

safety and product efficacy-safety and product efficacy-Change control ! Change control !

Validated Production Process

Δ


Less

More

Difficulty in Changing and Binding Authority

LAWS

Regulations

Guidance

Less

More

LAWS

Regulations

Guidance

Scientific Content & Detail about Implementation

Regulatory requirements Regulatory requirements for validationfor validation

21 CFR 211 Subpart H- Holding and Distribution21 CFR 211 Subpart H- Holding and Distribution– 211.165 – Testing and release for distribution211.165 – Testing and release for distribution

““Requires that the accuracy, sensitivity, Requires that the accuracy, sensitivity, specificity, and reproducibility of test methods specificity, and reproducibility of test methods employed by the firm shall be established and employed by the firm shall be established and documented. documented. Such validation and documentation Such validation and documentation may be accomplished in accordance with 21 CFR may be accomplished in accordance with 21 CFR 211.194 (a)(2)”211.194 (a)(2)”

Sec. 211.113 Control of microbiological contamination. Sec. 211.113 Control of microbiological contamination. (a) Appropriate written procedures, designed to prevent (a) Appropriate written procedures, designed to prevent

objectionable microorganisms in drug products not objectionable microorganisms in drug products not required to be sterile, shall be established and followed. required to be sterile, shall be established and followed.

(b) Appropriate written procedures, designed to prevent (b) Appropriate written procedures, designed to prevent microbiological contamination of drug products purporting microbiological contamination of drug products purporting to be sterile, shall be established and followed. to be sterile, shall be established and followed. Such Such procedures shall include validation of any sterilization procedures shall include validation of any sterilization process. process.

Regulatory requirements Regulatory requirements for validationfor validation

What does “validation of anyWhat does “validation of anysterilization process” mean ?sterilization process” mean ?

What parameters are critical to sterilization? What parameters are critical to sterilization?

– Temperatures, pressures, time, pore size (filtration), Temperatures, pressures, time, pore size (filtration), radiation dosage, chemical concentration.radiation dosage, chemical concentration.

Must demonstrate that your autoclave reaches the Must demonstrate that your autoclave reaches the temperatures, pressures, and times necessary for sterilization.temperatures, pressures, and times necessary for sterilization.

Must demonstrate that items representing real world samples Must demonstrate that items representing real world samples achieve those conditions ( 20 ft of 1 ½ hose; a 20 L carboy; a achieve those conditions ( 20 ft of 1 ½ hose; a 20 L carboy; a 500 ml bottle).500 ml bottle).

Must challenge with worse case scenario (may take place in Must challenge with worse case scenario (may take place in pilot plant if scalability demonstrated). pilot plant if scalability demonstrated).

Regulatory guidance on Regulatory guidance on validation validation

Guideline on General Principals of Process Validation Guideline 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 Documentation for Guidance for Industry: For the Submission Documentation for Sterilization Process Validation in Applications for Human and Sterilization Process Validation in Applications for Human and Veterinary Drug Products. CDER CVM November 1994. 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 Good Manufacturing to the EU Guide to Good Manufacturing

PracticePractice Title: Qualification and validationTitle: Qualification and validation http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdfhttp://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf ICH Q7a Section 12 on validation ICH 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. World Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World

Health Organization, Geneva. Health Organization, Geneva.

Critical Operations in Critical Operations in BiomanufacturingBiomanufacturing

Some operations are more critical than others. Some operations are more critical than others. – Viral filtration, sterilization, cleaning, analytical Viral filtration, sterilization, cleaning, analytical

methods. methods. – These operations will require greater validation These operations will require greater validation

efforts then less critical operations (media blending). efforts then less critical operations (media blending).

TestingTesting

Usually done by the Quality Control LaboratoryUsually done by the Quality Control Laboratory

– CFR requires that CFR requires that quality unitquality unit be under be under independent supervision and report directly to independent supervision and report directly to senior managementsenior management

Quality Quality AssuranceAssurance

Reviews records from quality control and production Reviews records from quality control and production departmentsdepartments– Verifies that all specifications and production Verifies 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

Designing Quality into Designing Quality into the Productthe Product

Design of production process and specifications Design 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 BSC for Clean rooms, closed systems, use of BSC for critical operations.critical operations.

– PurityPurity Separation process (chromatography) Separation process (chromatography)

designed to remove potential contaminantsdesigned to remove potential contaminants Viral purification / inactivationViral purification / inactivation

Validation Validation PlanPlan

Organizations must define an Organizations must define an approach towards validationapproach towards validation

– What is to be validated What is to be validated – How is it to be validatedHow is it to be validated– Who is to validate it Who is to validate it – Who is to approve the validationWho is to approve the validation– When it must be revalidated When it must be revalidated

Validation Validation Examples of individual Examples of individual

systems subject to systems subject to validation:validation:

HVAC systemsHVAC systemsAutoclavesAutoclavespH meterspH meters

Depyrogenation OvensDepyrogenation OvensLyopholyzersLyopholyzersCentrifugesCentrifuges

Steam generatorsSteam generatorsWater systemsWater systems

Compressed air systemsCompressed air systemsVacuum systemsVacuum systems

Validation PlanValidation Plan Regulatory agencies (FDA, EMEA, WHO, etc) identify Regulatory agencies (FDA, EMEA, WHO, etc) identify

minimum components of validation.minimum components of validation.

““Industry standards” (the c in cGMP) can increase Industry standards” (the c in cGMP) can increase validation requirements.validation requirements.

New & Novel processes / equipment require greater New & Novel processes / equipment require greater scrutiny then established processes / equipment. scrutiny then established processes / equipment.

Validation requirements increase as a product moves Validation requirements increase as a product moves through development (phase I, phase II, phase III).through development (phase I, phase II, phase III).

Validation PlansValidation PlansThe Validation Master Plan The Validation Master Plan

– A high level document that outlines the organizations A high level document that outlines the organizations philosophical approach to validation and revalidation. philosophical approach to validation and revalidation. The master validation plan becomes a guideline by The master validation plan becomes a guideline by which individual validation protocol are developed and which individual validation protocol are developed and implemented. implemented.

– May contain a flow chart or other diagram of the May contain a flow chart or other diagram of the validation processvalidation process

A prospective A prospective validation studyvalidation study

IQ

OQ

Calibration

PQ protocol approval

PQ protocol execution

Data Analysis

Validation Report

Approve Conclusions

Developmental Developmental studiesstudies

Experiments designed to explore and define the limits of Experiments designed to explore and define the limits of the system to be validated the system to be validated – Sterilization developmental studies may focus on Sterilization 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 “worst Cleaning developmental studies may focus on “worst

case” or hard to clean itemscase” or hard to clean items– Analytical methods may focus on defining the limits Analytical methods may focus on defining the limits

of the procedure (range, recovery, etc)of the procedure (range, recovery, etc)– Developmental studies then used to develop Developmental studies then used to develop

validation protocols and refine SOP’svalidation protocols and refine SOP’s

Validation ProtocolValidation Protocol Specific protocols (SOP’s) that provide detailed information Specific protocols (SOP’s) that provide detailed information

on what is to be validated.on what is to be validated.

Validation Protocols consist of: Validation Protocols consist of: – A description of the process, equipment, or method to be A description of the process, equipment, or method to be

validated.validated.– A description of the validation method.A description of the validation method.– A description of the sampling procedure including the A 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.

Example of a protocol for the IQ component of validating apH meter

As with all other SOP’s this document will contain an Objective, scope, and responsibilitySection.

Validation ProtocolValidation Protocol Validation Protocols may consist of multiple SOP’s each Validation Protocols may consist of multiple SOP’s each

describing specific steps in the validation processdescribing specific steps in the validation process

Critical SystemsCritical Systems How critical is the system being validated to final product How critical is the system being validated to final product

quality? quality?

– Media blending systems for cell growth vs. final fill & finish Media blending systems for cell growth vs. final fill & finish operationsoperations

Demonstrating that the device which fills, labels, and caps Demonstrating that the device which fills, labels, and caps the final product will require more extensive validation the final product will require more extensive validation then the blenders used to prepare media for bioreactors.then the blenders used to prepare media for bioreactors.

Validation of complex devices may take years! Validation of complex devices may take years!

ValidationValidation Proceeds in stages with new facilities / Proceeds in stages with new facilities /

equipment.equipment.

Planning for validation should start with the Planning for validation should start with the design process.design process.

Leaving validation to the last minute is Leaving validation to the last minute is asking for trouble. asking for trouble.

Stages of ValidationStages of Validation Starts with Design & Receipt:Starts with Design & Receipt:

– Does the equipment meet the needs (is the autoclave Does the equipment meet the needs (is the autoclave big enough?)big enough?)

– Do you have the manuals, spare parts, can you plug it Do you have the manuals, spare parts, can you plug it in? 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. and Does the autoclave reach the necessary temp. and pressure? pressure?

– Can the autoclave sterilize your equipment (worse Can the autoclave sterilize your equipment (worse case situation)?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?

IQ, OQ, PQ ? IQ, OQ, PQ ? Installation Qualification (IQ)Installation Qualification (IQ)

A process used to document that the piece of equipment was A process used to document that the piece of equipment was supplied and installed properly and that appropriate utilities, supplied and installed properly and that appropriate utilities, i.e., electrical, steam, gas, etc. are available to operate the i.e., electrical, steam, gas, etc. are available to operate the equipment according to the manufacturers specifications. equipment according to the manufacturers specifications.

Operational Qualification (OQ)Operational Qualification (OQ)A process designed to supply the documented evidence that a A process designed to supply the documented evidence that a piece of equipment operates as it is intended through all piece of equipment operates as it is intended through all anticipated operational ranges. anticipated operational ranges.

Performance (Process) Qualification (PQ)Performance (Process) Qualification (PQ)Verifies that a process / piece of equipment performs as it is Verifies that a process / piece of equipment performs as it is intended to in the manufacturing process and produces intended to in the manufacturing process and produces product (in process or final) meeting predetermined product (in process or final) meeting predetermined specifications. specifications.

Typical information in an Typical information in an IQ protocolIQ protocol

Name and description of equipment, including model Name 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, steam or Any utility requirements, i.e. electrical voltage, steam or

water pressure, etc.water pressure, etc. Any safety features of the equipment, including alarms, Any safety features of the equipment, including alarms,

interlocks, or relief valves.interlocks, or relief valves. That all documentation, including manufacturers contact That all documentation, including manufacturers contact

information, spare parts inventory, operational manual, and information, spare parts inventory, operational manual, and installation drawings are available on site. installation drawings are available on site.

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 an Typical information in an IQ protocolIQ protocol

Validation Validation 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 product however in some cases validation may take place as product is produced, or past production runs may be used to provide is produced, or past production runs may be used to provide validation data. validation data.

Prospective Validation Prospective Validation

Concurrent ValidationConcurrent Validation

Retrospective Validation Retrospective Validation

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 on the Periodic revalidation may be necessary depending on the

criticality of the equipmentcriticality of the equipment– Changes need to be evaluated for their impact on validationChanges need to be evaluated for their impact on validation– Deviations from specifications may require revalidationDeviations from specifications may require revalidation– Revalidation spelled out in Master Validation PlanRevalidation spelled out in Master Validation Plan

Change ControlChange Control Must assess impact of changes on FDA compliance and Must assess impact of changes on FDA compliance and

validation state. validation state.

Change control is a formal process defined in company Change control is a formal process defined in company SOP on how process/equipment changes are evaluated. SOP on how process/equipment changes are evaluated.

Any change that takes place outside the change control Any change that takes place outside the change control process can jeopardize product quality (patient safety).process can jeopardize product quality (patient safety).

Autoclave Validation Autoclave Validation IQ-IQ-

– Design specifications meet users needsDesign specifications meet users needs– Proper installation, utilities, manuals, Proper installation, utilities, manuals,

spare partsspare parts

The DQ, IQ, OQ process insures that this autoclave will meet the needs of the manufacturing group.

Sample Format for InstallationQualification 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.

Autoclave ValidationAutoclave Validation OQOQ

– Does it operate properlyDoes it operate properly Does it reach the specified temperature Does 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

Sample Format for OperationalQualification 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.

CalibrationCalibrationFigure 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.

Monitoring TemperatureMonitoring TemperatureFigure 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.

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.

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.

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.

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.

Developmental studiesDevelopmental studies Experiments designed to explore and define the Experiments designed to explore and define the

limits of the system to be validated limits of the system to be validated – Sterilization developmental studies may focus Sterilization developmental studies may focus

on “worst case ” or hard to sterilize items on “worst case ” or hard to sterilize items – Cleaning developmental studies may focus on Cleaning 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 the Analytical 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 develop Developmental studies then used to develop

validation protocols and refine SOP’svalidation protocols and refine SOP’s

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


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!


Does bagging make a difference?

Cleaning Validation Cleaning 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

How do we validate a How do we validate a dishwasher ? dishwasher ?

How do we identify hard to clean & easy to How 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?

Going over the documentation

Cleaning ValidationCleaning Validation

Sampling, documenting, and verifying is a labor intensive process

Why is cleaning considered a critical process?

SamplingSamplingSwabbing 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.

Cleaning Cleaning 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.

Analytical methods validationAnalytical methods validation Considered a critical step in the manufacturing process Considered a critical step in the manufacturing process

– Requirements for validated analytical methods Requirements for validated analytical methods explicitly written into the CFR’s explicitly written into the CFR’s

– 211.165 – Testing and release for distribution211.165 – Testing and release for distribution ““Requires that the accuracy, sensitivity, specificity, and Requires that the accuracy, sensitivity, specificity, and

reproducibility of test methods employed by the firm shall reproducibility of test methods employed by the firm shall be established and documented. Such validation and be established and documented. Such validation and documentation may be accomplished in accordance with documentation may be accomplished in accordance with 21 CFR 211.194 (a)(2)”21 CFR 211.194 (a)(2)”

Testing For IdentityTesting For Identity Requires the development of validated analytical Requires 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 to be?Is the molecule chemically what it is supposed to be? Biological Activity Tests:Biological Activity Tests:

– Does the molecules have the Does the molecules have the biologic activitybiologic activity that it is that it is supposed to have?supposed to have?

Immunogenic Tests:Immunogenic Tests:– Is the molecule immunogenic (allergic)? Is the molecule immunogenic (allergic)?

IdentityIdentity 21 CFR requires testing of raw materials:21 CFR requires testing of raw materials:

– Raw materials quarantined until identity Raw materials quarantined until identity verifiedverified

– Raw materials must meet Raw materials must meet predetermined predetermined specificationsspecifications

– Vendors (and alternates) specified in BLA (NDA)Vendors (and alternates) specified in BLA (NDA)

IdentityIdentity 21 CFR requires testing of in-process 21 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 - stop theMust meet specifications, if not - stop theprocess to investigate take corrective actionprocess to investigate take corrective action

Regulatory guidance on Regulatory guidance on validation validation

Guideline on General Principals of Process Validation Guideline 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 Documentation for Guidance for Industry: For the Submission Documentation for Sterilization Process Validation in Applications for Human and Sterilization Process Validation in Applications for Human and Veterinary Drug Products. CDER CVM November 1994. 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 Good Manufacturing to the EU Guide to Good Manufacturing

PracticePractice Title: Qualification and validationTitle: Qualification and validation http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdfhttp://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf ICH Q7a Section 12 on validation ICH 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. World Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World

Health Organization, Geneva. Health Organization, Geneva.

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

Analytical Methods ValidationAnalytical Methods Validation Being a critical component of production Being a critical component of production

process analytical methods must be process analytical methods must be validatedvalidated– Raw material testingRaw material testing– In process materialsIn process materials– Final product specifications Final product specifications

What must be What must be demonstrateddemonstrated

Selectivity (specificity)Selectivity (specificity) AccuracyAccuracy PrecisionPrecision Linear RangeLinear Range Limit of detection (LOD)Limit of detection (LOD) Limit of quantification (LOQ or Limit of quantification (LOQ or

LLOQ)LLOQ) RobustnessRobustness

Validated Validated methodsmethods

USP NF (United States Pharmacopeia National USP NF (United States Pharmacopeia National Formulary) contains “validated analytical methods” Formulary) contains “validated analytical methods”

Use of a USP method does not eliminate the Use of a USP method does not eliminate the organizations obligation to demonstrate that the method organizations obligation to demonstrate that the method performs adequetlyperforms adequetly

Selectivity Selectivity (specificity)(specificity)

Does the analytical method detect the component it is supposed Does the analytical method detect the component it is supposed to detect?to detect?– Cross reactivity in antibody based methods Cross reactivity in antibody based methods

Demonstrate specificity by conducting analytical method on Demonstrate 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”

AccurAccuracyacy

Ability of analytical method to accurately determine Ability 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 Typically done by analyzing a traceable

standardstandard

RecovRecoveryery

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” samples Typical recovery studies done using “spiked” samples Final results may be corrected by the % recovery Final results may be corrected by the % recovery

– Swab samples typically corrected to reflect recovery Swab samples typically corrected to reflect recovery

Linear Linear RangeRange

Must define the linear range of a method Must define the linear range of a method – Assay may have multiple linear rangesAssay may have multiple linear ranges

PrecisioPrecisionn

How much variability does the assay exhibit when analyzing the How much variability does the assay exhibit when analyzing the same samplesame sample– Typically demonstrated by analyzing multiple aliquots of a Typically demonstrated by analyzing multiple aliquots of a

homogenous samplehomogenous sample– Acceptance criteria will depend on the assay and the Acceptance 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)

Limit of Detection Limit of Detection (LOD)(LOD)

Lowest level at which method can detect Lowest level at which method can detect analyte analyte – Results reported as less than LOD Results reported as less than LOD – Based on signal to noise specification (10:1, Based on signal to noise specification (10:1,

20:1)20:1)

Robustness Robustness 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’s Detailed robustness studies will be reflected in final SOP’s

Limit of Limit 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 related Precision and LOQ related – Lower LOQ will typically result in lower precisionLower LOQ will typically result in lower precision

Bradford Bradford AssayAssay

for total for total proteinprotein

Well known colorimetric assay that relies on the binding of Well 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 in Dye 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

Bradford Bradford AssayAssay

DisadvantaDisadvantagesges

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

Validating the Validating 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

Some QuestionsSome Questions A valve used to transfer material from a holding tank to the A valve used to transfer material from a holding tank to the

purification suite jam’s closed. You have a spare valve that is an purification suite jam’s closed. You have a spare valve that is an identical model. Can you change this valve with the spare and identical model. Can you change this valve with the spare and continue operations? What if the valve is from a different continue operations? What if the valve is from a different manufacturer? manufacturer?

You notice that your autoclave loading plan leaves room for additional You notice that your autoclave loading plan leaves room for additional material. Realizing that increasing that amount of material in the material. Realizing that increasing that amount of material in the autoclave will shorten the turn around time for the production line you autoclave will shorten the turn around time for the production line you contemplate increasing the amount of material loaded into the contemplate increasing the amount of material loaded into the autoclave then specified by the loading plan. What should you do? autoclave then specified by the loading plan. What should you do? What will be required to implement this change? What will be required to implement this change?

An SOP for calibration of a pH meter calls for a two point calibration at An SOP for calibration of a pH meter calls for a two point calibration at pH 4 and pH 7. You notice that a single point calibration at pH 7 pH 4 and pH 7. You notice that a single point calibration at pH 7 produces the same result from pH measurements of your buffer produces the same result from pH measurements of your buffer solutions and allows you to take a longer break. Is it Ok to do the one solutions and allows you to take a longer break. Is it Ok to do the one point calibration when the SOP calls for a two point calibration? How point calibration when the SOP calls for a two point calibration? How would you go about changing the SOP to allow for a one point would you go about changing the SOP to allow for a one point calibration? calibration?

What documents would provide information concerning the What documents would provide information concerning the make and model of a particular valve used to regulate the make and model of a particular valve used to regulate the transfer of material from a holding tank to the purification suite? transfer of material from a holding tank to the purification suite?

Your supervisor is concerned that the fermentation vessel is not Your supervisor is concerned that the fermentation vessel is not providing sufficient aeration of the culture to get optimal growth providing sufficient aeration of the culture to get optimal growth and suggests installing a different kind of baffle in the vessel. and suggests installing a different kind of baffle in the vessel. How would you demonstrate that this change has no effect on How would you demonstrate that this change has no effect on product quality? product quality?

ReferencesReferences Pharmaceutical Manufacturers Association’s (Pharmaceutical Research and Manufacturers of America) Pharmaceutical Manufacturers Association’s (Pharmaceutical Research and Manufacturers of America)

Validation Advisory Committee “Process Validation Concepts for Drug Products” Pharmaceutical Technology, Validation Advisory Committee “Process Validation Concepts for Drug Products” Pharmaceutical Technology, September 1985 p 82.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, Marcel Decker, 2003. Pharmaceutical Process Validation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, Marcel Decker, 2003.

ISBN 082470838-5ISBN 082470838-5 Validation of Pharmaceutical Processes: Sterile Products. 1998. 2nd Edition. Edited by Frederick J. Carlton and Validation of Pharmaceutical Processes: Sterile Products. 1998. 2nd Edition. Edited by Frederick J. Carlton and

James Agalloco. Marcel Decker, 1998. ISBN 0824793846.James Agalloco. Marcel Decker, 1998. ISBN 0824793846. Validation Standard Operating Procedures: A step by Step Guide for Achieving Compliance in the Validation 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. ISBN Pharmaceutical, Medical Device, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002. ISBN 1574443313.1574443313.

Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control From Manufacturer to Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control From Manufacturer to Consumer, Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258. Consumer, Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.

Voss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBN 0939459507. Voss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBN 0939459507. LeBlanc, D.A. 2000. Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRC Press. ISBN LeBlanc, D.A. 2000. Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRC Press. ISBN

1574911163. 1574911163. Cloud, P. 1998. Pharmaceutical Equipment Validation: The Ultimate Qualification Guidebook. CRC Press. ISBN Cloud, P. 1998. Pharmaceutical Equipment Validation: The Ultimate Qualification Guidebook. CRC Press. ISBN

1574910795. 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. Pharmaceutical Technology DeSain C, Sutton C. (1995). Process development that supports process validation. Pharmaceutical Technology

19 (Oct.): 130-136, 1995.19 (Oct.): 130-136, 1995. Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a

powder mixture. powder mixture. Drug Development and Industrial PharmacyDrug Development and Industrial Pharmacy 27(4): 297-307, 2001. 27(4): 297-307, 2001. Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP)

requirements Part 2: Validation . World Health Organization, Geneva. 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. Stand.Dev. Biol. Stand. 81:81: 103-7 103-7 Nathanson, N. and Langmuir, A.D. 1995Nathanson, N. and Langmuir, A.D. 1995. . The Cutter incident. Poliomyelitis following formaldehyde-inactivated The Cutter incident. Poliomyelitis following formaldehyde-inactivated

poliovirus vaccination in the United States during the Spring of 1955. II. Relationship of poliomyelitis to Cutter poliovirus vaccination in the United States during the Spring of 1955. II. Relationship of poliomyelitis to Cutter vaccine. 1963. vaccine. 1963. Am. J. Epidemiol.Am. J. Epidemiol. 142:109-40142:109-40. .

Basis Validation of Biomanufacturing Processes

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