Incorporating Validation Concepts into the Biotechnology Curriculum (or minding your P’s and Q’s)

1
Incorporating Validation Concepts into the
Biotechnology Curriculum (or minding your Ps
and Qs)

• Thomas Burkett, Ph.D.
• The Community College of Baltimore County
• tburkett_at_ccbcmd.edu

2
Aims

• Introduce the role of validation in
  biomanufacturing
• Explain basic validation concepts and terminology
• Provide an example of how validation is
  incorporated into the biotech. curriculum at CCBC
• Provide examples of validation exercises
• Provide examples of validation resources

3
Validation in Biomanufacturing

• Biomanufacturing is a complex process involving
  multiple unit operations many of which are
  critical to insuring patient safety and product
  efficacy

4
Block Flow Diagram of a typical Production Process
5
Mammalian Antibody Production Cell Culture
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Mammalian Process Flow- Upstream Diagram
Transfer to Purification Suite
rProtein A Dia 1 meterCV 236 LBed 30cm
Virus Inactivation
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Mammalian Antibody Production - Downstream
Processing
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Validation in Biomanufacturing

• 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.

9
Validation in Biomanufacturing

• 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.

10
Validation in Biomanufacturing

• Validation does not replace testing, but it does
  reduce the testing 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-clinical material much less stringent then
  for phase III clinical material.
• Critical operations 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 and product efficacy

Validated Production Process
?
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Process Flow Diagram
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Regulatory requirement for validation

• 21 CFR 211 Subpart F Production and Process
  Controls
• 211.100 Written procedures deviations
• (a) Requires written procedures for production
  and process control designed to assure that
  products possess the quality attributes that they
  purport or are represented to possess.
• (b) Requires that any deviations from written
  production and process control procedures be
  recorded and justified.
• 211.101 Change in of components
• 211.103 Calculation of yield
• 211.105 Equipment identification
• 211.110 Sampling and testing of in-process
  materials and drug products
• Requires that control procedures be established
  to monitor the output and validate the
  performance of those manufacturing processes that
  may be responsible for causing variability of in
  process material and drug product.
• 211.111 Time limit on production
• 211.113 Control of microbiological
  contamination
• Requires that sterilization processes be
  validated
• 211.115 Reprocessing
• 21 CFR 211 Subpart H- Holding and Distribution
• 211.165 Testing and release for distribution
• Requires that the accuracy, sensitivity,
  specificity, and reproducibility of test methods
  employed by the firm shall be established and
  documented. Such validation and documentation may
  be accomplished in accordance with 21 CFR 211.194
  (a)(2)
• 21 CFR 211 Subpart I- Laboratory Controls
• 21 CFR 211 Subpart J Record and Reports
• 21 CFR 820 Quality Systems Regulations

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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 be sterile, shall be
  established and followed.
• (b) Appropriate written procedures, designed to
  prevent microbiological contamination of drug
  products purporting to be sterile, shall be
  established and followed. Such procedures shall
  include 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), radiation dosage, chemical
  concentration.
• Must demonstrate that your autoclave reaches the
  temperatures, pressures, and times necessary for
  sterilization.
• Must demonstrate that items representing real
  world samples achieve those conditions ( 20 ft of
  1 ½ hose a 20 L carboy a 500 ml bottle).
• Must challenge with worse case scenario (may take
  place in pilot plant if scalability
  demonstrated).

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FDA definition of validation

• Validation is a process of demonstrating,
  through documented evidence, that a process,
  procedure, method, piece of equipment, or
  facility will consistently produce a product or
  result that meets predetermined specifications
  and quality attributes.

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Regulatory guidance on validation

• Guideline on General Principals of Process
  Validation http//www.fda.gov/cder/guidance/pv.ht
  m
• Guidance for Industry For the Submission
  Documentation for Sterilization Process
  Validation in Applications for Human and
  Veterinary Drug Products. CDER CVM November 1994.
  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 Health Organization, 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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Validation in the biotech. curriculum at CCBC

• Validation introduced as part of quality
  systems section in intro. Course
• First lecture is on concepts of quality and
  quality attributes
• Second lecture introduces validation as part of
  the production process
• Lab exercises varies. Past examples include
  validation protocol for an autoclave validation
  of bioreactor sterilization.

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Learning Objectives

• Upon completion of this module students should
• Be familiar with the various government and third
  party literature pertaining to validation.
• Understand how component, process, and methods
  validation fits into the overall quality system.
• Be aware of pertinent regulations that apply to
  validation strategies.
• Understand concept of criticality and be able to
  identify points in the production process that
  are critical to product quality.
• Be able to distinguish between installation
  qualification, operation qualification, and
  performance qualification (IQ, OQ, PQ).
• Given the function of a piece of equipment used
  in biomanufacturing, discuss valtidation issues
  related to that specific piece of equipment.
• Be aware of the vendor, installation, and
  maintenance documentation required for initiating
  the validation process.
• Follow a validation SOP
• Be able to design a validation protocol for an
  individual piece of equipment.

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Concepts of Quality

• Biomanufacturing

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Quality Attributes

• Identity
• 21 CFR 211.84 (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 a product when prudently
  administered, taking into consideration the
  character of the product in relationship to the
  condition 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 at phase 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 by adequately controlled
  clinical data obtained through the administration
  of the product in the manner indicated to effect
  the given result.
• Purity
• 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.
• Cleaning Procedures
• Stability
• 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 may use accelerated time studies, biologics
  must use real time studies.
• Consistency

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Designing Quality into the product

• A central concept is that quality can not be
  tested for!
• Testing programs are based on testing a
  statistically significant number of samples
• However to be absolutely sure that all of your
  product meets specifications you would have to
  test everything.
• Testing by itself will not insure quality and is
  inefficient
• Testing is required under the GMPs
• Raw materials
• In-process samples
• Final Product
• Quality (identity, safety, efficacy, potency,
  purity, stability, consistency) must be designed
  into the production process
• Begins with predetermined specifications
• Raw material specifications
• In-process material specifications
• Final Product Specifications

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Predetermined specifications

• Identity
• Size, amino acid sequence, presence of post
  translational modifications, 3-D structure.

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Predetermined specifications
Identity
21 CFR 211.84 (d) at least one test shall be
conducted to verify the identity of each
component of a drug product. Tests consist of
Chemical, biological, and Immunological
methods. Requires testing of Raw materials,
In-process intermediates, final products.
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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 to have?
• 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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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 of BSC for
  critical operations.
• Purity
• Separation process (chromatography) designed to
  remove potential contaminants
• Viral purification / inactivation

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Insuring the Production of a Quality Product - II

• Validation its role in quality

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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 and assumptions!
• 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 ?

35
Validation

• The FDAs definition of validation
• Validation is a process of demonstrating,
  through documented evidence, that a process,
  procedure, method, piece of equipment, or
  facility will consistently produce a product or
  result that meets predetermined specifications
  and quality attributes.

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Quality Attributes Remember these?

• Identity
• 21 CFR 211.84 (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 a product when prudently
  administered, taking into consideration the
  character of the product in relationship to the
  condition 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 at phase 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 by adequately controlled
  clinical data obtained through the administration
  of the product in the manner indicated to effect
  the given result.
• Purity
• 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.
• Cleaning Procedures
• Stability
• 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 may use accelerated time studies, biologics
  must use real time studies.

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Historical Basis for Validation

• Assumptions concerning virus inactivation
  resulted in ten deaths and 200 children becoming
  paralyzed, from a supposedly inactivated polio
  vaccine.
• Assumptions about sterilization caused severe
  infections among burn victims given supposedly
  sterile solutions.
• Validation eliminates assumptions and relies on
  experimental proof!

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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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Validation Plan

• Regulatory agencies (FDA, EMEA, WHO, etc)
  identify minimum components of validation.
• Industry standards (the c in cGMP) can increase
  validation requirements.
• New Novel processes / equipment require greater
  scrutiny then established processes / equipment.
• Validation requirements increase as a product
  moves through development (phase I, phase II,
  phase III).

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Validation Plans

• The Validation Master Plan
• A high level document that outlines the
  organizations philosophical approach to
  validation and revalidation. The master
  validation plan becomes a guideline by which
  individual validation protocol are developed and
  implemented.
• May contain a flow chart or other diagram of the
  validation process

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Validation Protocol

• Specific protocols (SOPs) that provide detailed
  information on what is to be 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 and number 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.
43
Validation Protocol

• Validation Protocols may consist of multiple
  SOPs each describing specific steps in the
  validation process

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Validation

• Examples of individual systems subject to
  validation
• HVAC systems
• Autoclaves
• pH meters
• Depyrogenation Ovens
• Lyopholyzers
• Centrifuges
• Steam generators
• Water systems
• Compressed air systems
• Vacuum systems

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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 finish operations
• Demonstrating that the device which fills,
  labels, and caps the final product will require
  more extensive validation then the blenders used
  to prepare media for bioreactors.
• Validation of complex devices can 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 big enough?)
• 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
  case situation)?
• 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 and installed properly and
  that appropriate utilities, i.e., electrical,
  steam, gas, etc. are available to operate the
  equipment according to the manufacturers
  specifications.
• Operational Qualification (OQ)
• A process designed to supply the documented
  evidence that a piece of equipment operates as it
  is intended through all anticipated operational
  ranges.
• Performance (Process) Qualification (PQ)
• Verifies that a process / piece of equipment
  performs as it is intended to in the
  manufacturing process and produces product (in
  process or final) meeting predetermined
  specifications.

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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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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, and installation drawings are
  available on site.

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OQ Protocol
Example of a protocol for the OQ component of
validating a pH meter
As with all other SOPs this document will
contain an Objective, scope, and
responsibility Section.
52
OQ Protocol
Example of a protocol for the OQ component of
validating an autoclave
As with all other SOPs this document will
contain an Objective, scope, and
responsibility Section.
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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 product is produced, or past
  production runs may be used to provide validation
  data.
• Prospective Validation
• Concurrent Validation
• Retrospective Validation

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A prospective validation study
56
A concurent / retrospective validation study
No
Qualify system
Calibrate system
Yes
Yes
Approval
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The V-Model
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Planning for Validation

• Project Plan
• Agreed by team members
• Details phases, activities, and milestones
• Gantt Chart most commonly used

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• Putting it
• all together

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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 criticality of the equipment
• Changes need to be evaluated for their impact on
  validation
• Deviations from specifications may require
  revalidation
• Revalidation spelled out in Master Validation 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 how process/equipment changes are
  evaluated.
• Any change that takes place outside the change
  control process can jeopardize product quality
  (patient safety).

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An example of a facility / process validation

• Remicade (infliximab) is a chimeric mAb
  directed against TNF-a.
• Approved in 1998 (US) and 1999 (EU) to treat
  Crohns disease, and RA.
• Produced by Centocor, Inc. in Malvern, PA
• Contains mouse variable domains and human
  constant domains (IgG1)

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Antibodies

• Proteins
• 2 heavy Chains
• 2 Light Chains
• Disulfide Bonds
• Variable region
• Recognizes antigen
• Constant region
• Effecter function
• Classes subclasses

Ig G class
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Production of Remicade

• BLA approved in August 1998 (FDA), 1999 (EMEA).
• First site for bulk manufacture was Leiden, The
  Netherlands.
• Process was transferred to Malvern, PA in April
  2002.
• Process changes, including larger bioreactors,
  external spin filters, and a change in media
  components were introduced to meet increased
  demand.
• Not only did a new facility have to be validated,
  but also the changes to the manufacturing process
  had to be validated.
• Necessary to demonstrate that product produced
  under these new conditions had same quality
  attributes as product produced in Leiden.
• An unanticipated consequence of increased product
  yield was a change in chromatography conditions
  due to product breakthrough under old conditions.
• Minor changes can have unanticipated consequences
  on product quality!
• A new facility for production of remicade is
  being constructed in the Republic of Ireland and
  should be on line in 2007

65
Changes in Production Process in Malvern, PA
66
Example of a 1000 L Bioreactor with an external
spin filter used in the production of Remicade
in Malvern, PA
67
Remicade Production
68
These tanks are used for the holding of material
from the bioreactors prior to product capture
and initial chromatography. What performance
aspects of these tanks do you think need to be
validated? How does cleaning of these tanks
between use affect validation?
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Some Questions

• A valve used to transfer material from a holding
  tank to the purification suite jams closed. You
  have a spare valve that is an identical model.
  Can you change this valve with the spare and
  continue operations? What if the valve is from a
  different manufacturer?
• You notice that your autoclave loading plan
  leaves room for additional material. Realizing
  that increasing that amount of material in the
  autoclave will shorten the turn around time for
  the production line you contemplate increasing
  the amount 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 4 and 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 a
  longer break. Is it Ok to do the one 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 calibration?

70
Some More Questions

• 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?

71
References

• Pharmaceutical Manufacturers Associations
  (Pharmaceutical Research and Manufacturers of
  America) Validation Advisory Committee Process
  Validation Concepts for Drug Products
  Pharmaceutical Technology, September 1985 p 82.
• 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. ISBN 082470838-5
• Validation of Pharmaceutical Processes Sterile
  Products. 1998. 2nd Edition. Edited by Frederick
  J. Carlton and James Agalloco. Marcel Decker,
  1998. ISBN 0824793846.
• 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 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. Pharmaceutical Equipment
  Validation 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
  Technology 19 (Oct.) 130-136, 1995.
• Garcia T, Wilkinson S, Scott J. The development
  of a blend-sampling technique to assess the
  uniformity of a powder mixture. Drug Development
  and Industrial Pharmacy 27(4) 297-307, 2001. 
• 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.
• 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 poliovirus vaccination
  in the United States during the Spring of 1955.
  II. Relationship of poliomyelitis to Cutter
  vaccine. 1963. Am. J. Epidemiol. 142109-40.

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Laboratory Activities

• Students develop and carry out a simplified
  validation plan / protocol
• Autoclave validation
• Bioreactor sterilization
• Bioreactor cleaning
• Spectroscopy
• Chromatography
• Plasmid construct

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