validation 100908
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Validation & Biomanufacturing
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What is Validation
Validation An Essential Part of GMPs!
Validation is the scientific study of a system
To prove that the facility/system/equipment/method is consistently doing
what it is supposed to do (i.e., that the process is under control).
We want to make decisions based on good science and not hunches andassumptions!
To determine the process variables and acceptable limits for these variables,
and to set-up appropriate in-process controls.
Is it ok if the wash from a chromatography column is pH 6.8 vs. 7.0 ?
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FDA definition of validation
Validation is a process ofdemonstrating, through
documented evidence, that a process, procedure,method, piece of equipment, or facility will
consistentlyproduce a product or result that meets
predetermined specifications and quality
attributes.
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Quality Attributes Remember these?Identity
21 CFR211.84 (d) at least one test shall be conducted to verify the identity of each component of a drug product.
Chemical, biological, Immunological Raw materials, In-process intermediates, final products.
Safety
21 CFR 600.3 (p) safety as the relative freedom from harmful effect to persons affected, directly or indirectly, by aproduct when prudently administered, taking into consideration the character of the product in relationship to thecondition of the recipient at the time.
Activity of active ingredients
Activity of the excipients or additives
Activity of process related impurities
Efficacy
Effectiveness of the product in achieving its medicinal purpose (therapeutic, prophylactic, diagnostic). Gathered atphase II and Phase III trials.
Potency
21 CFR 600.3 (s) specific ability or capacity of the product, as indicated by its appropriate laboratory tests or byadequately controlled clinical data obtained through the administration of the product in the manner indicated toeffect the given result.
Purity
21 CFR 600.3 (r) relative freedom from extraneous matters in the finished product, whether or not harmful to therecipient or deleterious to the product.
Cleaning Procedures
Stability
21 CFR211.137 (a) to assure that a drug product meets applicable standards of identity, quality, and purity at thetime of use; it shall bear an expiration date determined by stability testing. Drugs may use accelerated time studies,biologics must use real time studies.
Consistency
The ability of the product and/or process to reliably possess specified quality attributes on an ongoing basis. 3consecutive batches of product meeting predetermined specifications is accepted as proof that a process isconsistent. However, in NDA data from up to twenty batches may be submitted.
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Biomanufacturing
Biomanufacturing is a complex process involving multiple unit
operations many of which are critical to insuring patient safety and
product efficacy
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InnoculumSeed
Fermentati
on
ProductionFermentati
on
Harvest
Ultrafiltrati
on
1
Chrom. 11
Ultrafiltrati
on
2
Chrom.
2
Viral
Filtration
Chrom.
3
Ultrafiltrati
on
3
Final
Formulation/
Sterile Filtration
Sterile Fill
UPSTREAM
DOWN-
STREAM
VIRAL
NON-VIRAL
Block Flow Diagram of a typical Production Process
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Historical Basis for Validation
Assumptions concerning virus inactivation resulted in tendeaths and 200 children becoming paralyzed, from asupposedly inactivated polio vaccine.
Assumptions about sterilization caused severe infectionsamong burn victims given supposedly sterile solutions.
Validation eliminates assumptions and relies on
experimental proof!
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Quality by Design
A central concept in quality is that quality can not be tested for.
Quality must be designed and built into the production process.
Requires careful attention to raw material specifications, in
process material specifications, and final product specifications.
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Validation and Quality
Validating the performance of unit operations, analytical
methods, and critical process points (sterilization, viral
inactivation, cleaning procedures) is essential in insuring that
the process generates a quality product.
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Validation in Biomanufacturing
Validation does not replace testing, but it does reduce thetesting burden for raw materials, in-process materials, and
final product
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Validation in Biomanufacturing
Validation itself is a process that evolves with the
product.
Validation requirements for production of pre-clinicalmaterial less stringent then for phase III clinical
material.
Critical operations must be validated: For example:
raw materials, analytical methods, viral clearance,sterilization, cleaning.
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Validation in Biomanufacturing
A fully validated process is locked in
Any change outside of the validated space invalidates process
Change must be evaluated for effect on patient safety andproduct efficacy-Change control !
Validated Production Process
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Less
More
Difficulty in Changing and Binding Authority
LAWS
Regulation
Guidance
Less
More
LAWS
Regulation
Guidance
Scientific Content & Detail about Implementation
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Regulatory requirement for validation
21 CFR211 Subpart H- Holding and Distribution
211.165 Testing and release for distribution
Requires that the accuracy, sensitivity, specificity, andreproducibility of test methods employed by the firmshall be established and documented. Such validationand documentation may be accomplished in accordance
with 21 CFR 211.194 (a)(2)
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Regulatory requirement for validation
Sec. 211.113 Control of microbiological contamination.
(a) Appropriate written procedures, designed to prevent
objectionable microorganisms in drug products not required to besterile, shall be established and followed.
(b) Appropriate written procedures, designed to preventmicrobiological contamination of drug products purporting to besterile, shall be established and followed. Such procedures shallinclude validation of any sterilization process.
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What does validation of any
sterilization process mean ? What parameters are critical to sterilization?
Temperatures, pressures, time, pore size (filtration), radiationdosage, chemical concentration.
Must demonstrate that your autoclave reaches the temperatures,pressures, and times necessary for sterilization.
Must demonstrate that items representing real world samples achievethose conditions ( 20 ft of1 hose; a 20 L carboy; a 500 ml bottle).
Must challenge with worse case scenario (may take place in pilotplant if scalability demonstrated).
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Regulatory guidance on validation
Guideline on General Principals of Process Validationhttp://www.fda.gov/cder/guidance/pv.htm
Guidance for Industry: For the Submission Documentation for SterilizationProcess Validation in Applications for Human and Veterinary DrugProducts. CDER CVM November1994.www.fda.gov/CDER/GUIDANCE/cmc2.pdf
Working Party on Control of Medicines and Inspections Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice
Title: Qualification and validation
http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf
ICH Q7a Section 12 on validation
http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm
A WHO guide to good manufacturing practice (GMP) requirements. Part 2:Validation
Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World HealthOrganization, Geneva.
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Critical Operations in
Biomanufacturing
Some operations are more critical than others.
Viral filtration, sterilization, cleaning, analytical methods.
These operations will require greater validation efforts then less
critical operations (media blending).
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Testing
Usually done by the Quality Control Laboratory
CFR requires that quality unit be under independent
supervision and report directly to senior management
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Quality Assurance
Reviews records from quality control and production departments
Verifies that all specifications and production operations met /performed
Investigations necessary for any deviations
Root cause
Affect on quality
Corrective action (CAPA)
Approves final release of product
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Designing Quality into the Product Design of production process and specifications all contribute
to a quality product:
Absence of contamination
Clean rooms, closed systems, use ofBSC for criticaloperations.
Purity
Separation process (chromatography) designed toremove potential contaminants
Viral purification / inactivation
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Validation Plan
Organizations must define an approach towards
validation
What is to be validated
How is it to be validated
Who is to validate it
Who is to approve the validation When it must be revalidated
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ValidationExamples of individual
systems subject to validation:
HVAC systems
Autoclaves
pH meters
Depyrogenation Ovens
Lyopholyzers
Centrifuges
Steam generatorsWater systems
Compressed air systems
Vacuum systems
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Validation Plan
Regulatory agencies (FDA, EMEA, WHO, etc) identify minimumcomponents of validation.
Industry standards (the c in cGMP) can increase validationrequirements.
New & Novel processes / equipment require greater scrutiny thenestablished processes / equipment.
Validation requirements increase as a product moves throughdevelopment (phase I, phase II, phase III).
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A prospective validation study
IQ
OQ
Calibration
PQ protocol
approval
PQ protocolexecution
Data
Analysis
ValidationReport
Approve
Conclusions
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Developmental studies
Experiments designed to explore and define the limits of the system to be
validated
Sterilization developmental studies may focus on
worst case orhard to sterilize items
Cleaning developmental studies may focus on worst case or hard to
clean items
Analytical methods may focus on defining the limits of the procedure
(range, recovery, etc)
Developmental studies then used to develop validation protocols and
refine SOPs
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Validation Protocol
Specific protocols (SOPs) that provide detailed information on what is tobe validated.
Validation Protocols consist of: A description of the process, equipment, or method to be validated.
A description of the validation method.
A description of the sampling procedure including the kind andnumber of samples.
Acceptance criteria for test results. Schedule or criteria for revalidation.
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Example of a protocol for the
IQ component of validating a
pH meter
As with all other SOPs this
document will contain an
Objective, scope, and responsibility
Section.
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Validation Protocol
Validation Protocols may consist of multiple SOPs each
describing specific steps in the validation process
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Critical Systems
How critical is the system being validated to final product quality?
Media blending systems for cell growth vs. final fill & finishoperations
Demonstrating that the device which fills, labels, and caps thefinal product will require more extensive validation then the
blenders used to prepare media for bioreactors.
Validation of complex devices may take years!
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Validation
Proceeds in stages with new facilities / equipment.
Planning for validation should start with the design
process.
Leaving validation to the last minute is asking for
trouble.
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Stages of Validation
Starts with Design & Receipt:
Does the equipment meet the needs (is the autoclave bigenough?)
Do you have the manuals, spare parts, can you plug it in?
Is it installed properly (drain lines, vents, etc) Does it work?
Does the autoclave reach the necessary temp. and pressure?
Can the autoclave sterilize your equipment (worse casesituation)?
How does it work in the manufacturing process? Can it handle production quantities?
Will failure compromise product quality?
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IQ, OQ, PQ ?
Installation Qualification (IQ)
A process used to document that the piece of equipment was supplied andinstalled properly and that appropriate utilities, i.e., electrical, steam, gas,etc. are available to operate the equipment according to the manufacturersspecifications.
Operational Qualification (OQ)
A process designed to supply the documented evidence that a piece ofequipment operates as it is intended through all anticipated operationalranges.
Performance (Process) Qualification (PQ)Verifies that a process / piece of equipment performs as it is intended to inthe manufacturing process and produces product (in process or final)meeting predetermined specifications.
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Typical information in an IQ protocol
Name and description of equipment, including model numbers
Identification, including model and serial numbers
Location of the equipment Any utility requirements, i.e. electrical voltage, steam or water
pressure, etc.
Any safety features of the equipment, including alarms, interlocks,or relief valves.
That all documentation, including manufacturers contact
information, spare parts inventory, operational manual, andinstallation drawings are available on site.
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Typical OQ Protocol Components
Objective
Responsibility Equipment required (Calibration verification & Traceability)
SOP(s) used
Equipment Identification
Parameters measured (Specifications)
Documentation
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Validation
Ideally validation takes place prior to actual production runs,however in some cases validation may take place as productis produced, or past production runs may be used to providevalidation data.
ProspectiveValidation
ConcurrentValidation
Retrospective Validation
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Revalidation
Is the initial validation of a piece of equipment the end?
No!
Periodic revalidation may be necessary depending on the criticalityof the equipment
Changes need to be evaluated for their impact on validation
Deviations from specifications may require revalidation
Revalidation spelled out in MasterValidation Plan
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Change Control
Must assess impact of changes on FDA compliance and validation
state.
Change control is a formal process defined in company SOP on howprocess/equipment changes are evaluated.
Any change that takes place outside the change control process canjeopardize product quality (patient safety).
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References Pharmaceutical Manufacturers Associations (Pharmaceutical Research and Manufacturers of America) Validation
Advisory Committee ProcessValidation Concepts for Drug Products Pharmaceutical Technology, September1985 p 82.
Bismuth, G. CleaningValidation: A Practical Approach. CRC Press, 2000. ISBN 1574911082. Pharmaceutical ProcessValidation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, Marcel Decker, 2003. ISBN
082470838-5
Validation of Pharmaceutical Processes:Sterile Products. 1998. 2nd Edition.Edited by Frederick J. Carlton and JamesAgalloco. Marcel Decker, 1998. ISBN 0824793846.
Validation Standard Operating Procedures:A step by Step Guide for Achieving Compliance in the Pharmaceutical, MedicalDevice, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002. ISBN 1574443313.
Good Manufacturing Practices for Pharmaceuticals:A Plan for Total Quality Control From Manufacturer to Consumer,Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.
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 1574911163. Cloud, P. 1998. PharmaceuticalEquipmentValidation: The Ultimate Qualification Guidebook. CRC Press. ISBN
1574910795.
Juran, Quality Control Handbook, 4th Edition., McGraw-Hill, 1988.
DeSain C, Sutton C. (1995). Process development that supports process validation. Pharmaceutical Technology19 (Oct.):130-136, 1995.
Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a powdermixture. DrugDevelopment and Industrial Pharmacy27(4):297-307,2001.
Chaloner-Larsson, G., Anderson, R.,Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirementsPart 2:Validation . World Health Organization, Geneva. www.who.int/vaccines-documents/DocsPDF/www9666.pdf
Accessed on October2nd, 2006. Brown, F. 1993. Review of accidents caused by incomplete inactivation of viruses. Dev. Biol. Stand. 81: 103-7
Nathanson, N. and Langmuir, A.D. 1995. The Cutter incident. Poliomyelitis following formaldehyde-inactivated poliovirusvaccination in the United States during the Spring of1955. II. Relationship of poliomyelitis to Cutter vaccine. 1963.Am. J.Epidemiol.142:109-40.
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Autoclave Validation
IQ-
Design specifications meet users needs
Proper installation, utilities, manuals, spare parts
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The DQ, IQ, OQ process
insures that this autoclave
will meet the needs of the
manufacturing group.
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Sample Format forInstallationQualification 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.
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Autoclave Validation
OQ
Does it operate properly Does it reach the specified temperature and
pressure
Do timers work
Does the operator interface panel work
Are safety interlocks functional
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Sample Format forOperational
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.pdfAccessed on October 2nd, 2006.
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Monitoring Temperature
Figure 2: A validator, used in the
operational qualification of an
autoclave. The validator is
attached to individual
thermocouples by wires comingfrom 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 thecomputer is also subject to
validation requirements.
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Figure 4. The validator is
attached to individual
thermocouples (TC) by thin
wires that pass through the
wall of the autoclavethrough a specially
designed port (arrow). This
picture shows the back side
of the autoclave. The
validator is out of view at
the lower left.
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Figure 5. The inside of the
autoclave showing the maze
of wiring connecting the
individual TCs to the
validator. The port through
which the wires pass is
visible in the middle left of
the picture. The individual
TCs will be placed in various
areas of the autoclave or
equipment being autoclavedto generate a thermal map of
the interior of the autoclave.
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Figure 3. Output from the
validator. The temperature at
each connected thermocouple is
displayed. Accumulated lethality
(F0) may also be displayed.Notice how some TChave failed
and will not record a
temperature. Accounting for TC
failure is necessary to keep from
having to repeat a study.
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Developmental Studies
Developmental studies are
used to identify hard to
autoclave items and totest if item preparation has
an effect on ability to be
sterilized
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Developmental 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!
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Developmental Studies
Does bagging make a difference?
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Cleaning Validation
Validating the cleaning cycle
on a loaded dishwasher.
Notice the various pipes andparts with narrow openings.
Identification of hard to clean
and easy to clean areas starts
with developmental studies
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How do we validate a dishwasher ?
How do we identify hard to clean & easy to clean items?
How do we test to see if they are cleaned?
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Going over the documentation
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Cleaning Validation
Sampling, documenting, and
verifying is a labor intensive
process
Why is cleaning considered a
critical process?
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Sampling
Figure 8. Swabbing is
commonly used to sample the
surface of cleaned materials.
The swab is then placed in a
sample vial and sent to theQuality 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 knowndeficiencies in recovery.
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Cleaning Validation
Figure 9. Analysis of rinse water
for residual cleaning agents orprocess materials is an essential
component of cleaning validation.
Insuring that the sample is not
contaminated requires vigilance
and properly following the
relevantSOPs.
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Analytical methods validation
Considered a critical step in the manufacturing process
Requirements for validated analytical methods explicitly written intothe CFRs
211.165 Testing and release for distribution
Requires that the accuracy, sensitivity, specificity, and reproducibility oftest methods employed by the firm shall be established and documented.Such validation and documentation may be accomplished in accordancewith 21 CFR211.194 (a)(2)
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Testing For Identity
Requires the development of validated analytical methods that can
determine identity. Chemical Tests:
Is the molecule chemically what it is supposed to be?
Biological Activity Tests:
Does the molecules have the biologic activity that it is supposed tohave?
Immunogenic Tests:
Is the molecule immunogenic (allergic)?
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Identity
21 CFR requires testing of raw materials:
Raw materials quarantined until identity verified
Raw materials must meet predetermined specifications
Vendors (and alternates) specified in BLA (NDA)
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Identity
21 CFR requires testing of in-process materials:
Product from bioreactor / fermentor Product from purification steps
Waste products from above
Must meet specifications, if not - stop the
process to investigate take corrective action
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Regulatory guidance on validation
Guideline on General Principals of Process Validationhttp://www.fda.gov/cder/guidance/pv.htm
Guidance for Industry: For the Submission Documentation for SterilizationProcess Validation in Applications for Human and Veterinary DrugProducts. CDER CVM November1994.www.fda.gov/CDER/GUIDANCE/cmc2.pdf
Working Party on Control of Medicines and Inspections
Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice
Title: Qualification and validation
http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf
ICH Q7a Section 12 on validation
http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm
A WHO guide to good manufacturing practice (GMP) requirements. Part 2:Validation
Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World HealthOrganization, Geneva.
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Validation Protocol
Specific protocol based on developmental studies
Protocol is written, reviewed and approved Protocol is executed
Report written and approved
System is validated
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Analytical Methods Validation
B
eing a critical component of productionprocess analytical methods must be validated
Raw material testing
In process materials
Final product specifications
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What must be demonstrated
Selectivity (specificity)
Accuracy
Precision Linear Range
Limit of detection (LOD)
Limit of quantification (LOQ or LLOQ)
Robustness
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Validated methods
USP NF (United States Pharmacopeia National Formulary) contains
validated analytical methods
Use of a USP method does not eliminate the organizations
obligation to demonstrate that the method performs adequetly
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Selectivity (specificity)
Does the analytical method detect the component it is supposed
to detect? Cross reactivity in antibody based methods
Demonstrate specificity by conducting analytical method on
materials that may mimic analyte of interest
Looking forfalse positives
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Accuracy
Ability of analytical method to accurately determine the
presence and amount of the analyte of interest Typically done by analyzing a traceable standard
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Linear Range
Must define the linear range of a method
Assay may have multiple linear ranges
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Precision
How much variability does the assay exhibit when analyzingthe same sample
Typically demonstrated by analyzing multiple aliquots of a
homogenous sample
Acceptance criteria will depend on the assay and the
material being assayed (2-20% RSD)
Typically expressed as % RSD (relative standard deviation)
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Limit ofDetection (LOD)
Lowest level at which method can detect analyte
Results reported as less than LOD
Based on signal to noise specification (10:1, 20:1)
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Limit ofQuantification (LOQ)
Lowest level at which method can accurately quantify analyte
Based on signal-to-noise ratio specification (10:1, 20:1)
and precision specification
Precision and LOQ related
Lower LOQ will typically result in lower precision
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Bradford Assayfor total protein
Well known colorimetric assay that relies on the binding of
Commassie G-250 dye to the proteins in an acidic solution
Dye binding proportional to number of positive charges in
protein
Proteins >3000 dal not detected
Simple, quick, wide range, few interfering agents
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BradfordA
ssay
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Disadvantages
Incompatability with surfactants Staining of glass and quartz cuvettes
Use disposable polystyrene cuvettes
Or wash with strong detergents and methanol
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Validating the Bradford
Selectivity (specificity)
Accuracy
Recovery
Precision
Linear Range
Limit of detection (LOD)
Limit of quantification (LOQ or LLOQ) Robustness
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Some Questions
A valve used to transfer material from a holding tank to the purification suitejams closed. You have a spare valve that is an identical model. Can you changethis valve with the spare and continue operations? What if the valve is from adifferent manufacturer?
You notice that your autoclave loading plan leaves room for additional material.
Realizing that increasing that amount of material in the autoclave will shortenthe turn around time for the production line you contemplate increasing theamount of material loaded into the autoclave then specified by the loading plan.What should you do? What will be required to implement this change?
An SOP for calibration of a pH meter calls for a two point calibration at pH 4and pH 7. You notice that a single point calibration at pH 7 produces the same
result from pH measurements of your buffer solutions and allows you to take alonger break. Is it Ok to do the one point calibration when the SOP calls for atwo point calibration? How would you go about changing the SOP to allow for aone point calibration?
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What documents would provide information concerning the make and
model of a particular valve used to regulate the transfer of material from a
holding tank to the purification suite?
Your supervisor is concerned that the fermentation vessel is not providing
sufficient aeration of the culture to get optimal growth and suggests
installing a different kind of baffle in the vessel. How would you
demonstrate that this change has no effect on product quality?