BIO 221 BIOMANUFACTURING AND CGMP
COMPLIANCE
LABORATORY COURSE
LABORATORY
EXERCISE NUMBER
TITLE
1
DOCUMENTATION, QUALITY
SYSTEMS AND GMP (PART 1-SOP
EXERCISE)
2
DOCUMENTATION, QUALITY
SYSTEMS AND GMP (PART 2-BATCH
RECORD AUDIT)
3
DOCUMENTATION, QUALITY
SYSTEMS AND GMP (PART 3-BATCH
RECORDS)
4
DOCUMENTATION, QUALITY
SYSTEMS AND GMP (PART 4-BATCH
RECORDS, CONTINUED)
5
GOWNING
6
AUTOCLAVE MONITORING
7
GMP POPCORN
DOCUMENTATION, QUALITY SYSTEMS AND GMP
Background
Good documentation practices are an essential feature of any biotechnology company. A
document affirms the following and contains such a statement depending on the purpose
of the document: The rules set forth in this document should apply to all personnel in a
biotechnology company involved in the holding, transportation, manufacture, testing,
support, or packaging of clinical and commercial drugs, whether temporary or regular
staff.
Biotechnology companies are regulated by the Food and Drug Administration (FDA) as
well as by foreign regulatory agencies for products marketed abroad. Strict
documentation rules are set to ensure compliance with all regulatory agencies. By placing
your signature on a cGMP document, you have proven in the view of the FDA that you
have completed a step. cGMP documents and records include, but are not limited to the
following: Standard Operating Procedures (SOPs), Manufacturing Procedures (MPs),
Specifications, Analytical Methods, Validation Documents, Batch Records, Product and
Sample Labels. All cGMP documents may be reviewed by the FDA or other regulatory
agencies.
Documentation Rules
1. Do
When writing in cGMP documents DO:
 Use only black, indelible, ball-point ink
 Make all entries legible such that they are readable
 Initial, unless a signature is required, and date all entries
 Document each step before moving to the next
 Write N/A, initial and date spaces if it is not appropriate to fill them in
 Record numbers less than one with a zero before the decimal point
2. Do Not
When writing in cGMP documents DO NOT:
 Overwrite
 Use liquid correction fluid
 Backdate
 Record data before the action or event has occurred
 Use ditto marks
 Leave required data spaces blank
 Approve, verify or review your own performance
3. Initial/Date
All entries to a cGMP document must be accompanied by the identity of the person
(initials or signature) and the date that the entry was made. This is required by the Code
of Federal Regulations (CFR) and serves as a tracking method to determine that a task
was indeed performed and who did the work.
Initials are the accepted standard method of identification. However, some operations
require a signature. For example, an “Approved By” space must be filled with a
signature, not initials.
4. Recording Time
Either military time or meridian time is acceptable for recording time.
Military time: two (2) digits to indicate the hour (00 to 23) followed by two (2) digits to
indicate the minutes (00 to 59).
Examples:
0850
1750
Meridian time: One or two digits to indicate the hour (1 to 12) followed by two digits to
indicate the minutes (00 to 59), then the morning (AM) or afternoon (PM) designation.
Examples:
8:50 AM
5:50 PM
5. Corrections
No handwritten changes or corrections may be made to the printed text of an approved
cGMP document. Consult with your supervisor if you discover an error. Any changes
required to an approved cGMP document shall be implemented through the established
quality system.
When making a correction to a manually recorded entry on controlled documents perform
the following steps:
 Place a single line through the incorrect entry
 Initial and date adjacent to the cross-out
 Enter the correct data near the original entry
 The mistake must still be legible through the cross-out
 Date of correction is the date the correction was made, not the date the error was
made.
6. Performed By
Performance of a step must be documented at the time of completing the step and prior to
moving on to the next step. Do not execute a step if the manufacturing procedure is not
available for documenting necessary data at the time of execution. The following
personnel may initial and date the “Performed By” space:
 Personnel already proficient in the task performed OR

Personnel who are in training under the supervision of their qualified trainer
7. Recorded By
The “Recorded By” space is used if the operator is unable to initial and date immediately,
due to working in a confined or restricted space, such as a laminar air flow hood. This
situation is the only exception to the “Performed By” rule. Data must then be recorded by
another person watching the operation. The person recording data must initial and date
the “Recorded By” space prior to moving on to the next step.
8. Verified By
Verification shall be performed prior to moving on to the next step. Operators executing a
task cannot verify their own action. At least one other person must review documentation
for accuracy. Personnel may initial or sign and date the “Verified By” space if:
 They witnessed that a task, operation, or procedure was performed per written
instructions and accurately documented AND
 They are already proficient in the task performed
9. Deviations
If you deviate from a written procedure, you must:
 Notify your supervisor
 Document the deviation using the appropriate quality system
10. Missing Data
If information is not entered at the time of completing the step, the blank entry shall be
marked by an asterisk or similar notation. The use of each notation is limited to one per
page. Comments explaining the reason information is missing, along with the proper
information, e.g. date event actually occurred, shall be documented on the same page of
the record. The explanation shall be initialed and dated at the time of recording.
11. Voiding Records
On occasion, errors are made in the execution of making an in-process material such as a
buffer and all the proper documentation was completed. However, because of the error,
the decision is made to scrap the material and start all over with new in-process material.
The original document would then need to be voided and attached to the document
replacing it. The documents are voided to prevent confusion or mix-ups with the correct
document. When voiding a document do the following:
 Get supervisor and Quality Assurance (QA) approval
 Write “Void” across the front of the document and include initials and date
12. Recreating and Rewriting Records
Recreating or rewriting records should be avoided; however it is sometimes necessary.
Supervisor and QA approval is required when recreating records. It is important to
identify the recreated document as “Rewrite” and to reference the sources of the
information. Records can be recreated only when:
 Record is illegible
 Incorrect form or document was used
 Record was irreparably damaged
13. Rounding Off Rules
The following rules apply to rounding off:
 In a series of calculations, carry the extra digits through the final result, and then
round off.
 If the digit to be removed is <5, the preceding digit remains the same. For
example, 1.84 rounds to 1.8.
 If the digit to be removed is ≥5, the preceding digit is increased by 1. For
example, 1.85 rounds to 1.9.
GLOSSARY
Approved By
Backdating
Batch
Production
Record
Comment
Controlled
Documents
Cross-out
Data
Date
Document
Signature of a qualified individual (supervisor or designee)
indicating that the information documented is complete,
accurate, and acceptable.
Backdating is the practice of going back to a previously
completed task that has not been properly initialed and dated and
placing the date that the task was completed on the date line, as
though filling in the date had been done in a timely fashion. This
practice is not allowed in any cGMP document.
Collection of records associated with the manufacture of a
specific lot of product.
Any written additions to a document for informational purposes.
All comments must be initialed and dated by the person writing
the addition and may require a verification.
Written approved documents used in association with cGMPrelated activities to ensure compliance with US and international
regulations, as well as company standards.
A cross-out indicates a correction has been made. This is
accomplished by drawing a single straight ink line through
information which has been entered inadvertently or incorrectly.
The values and information generated by processing, calculating
or transcribing from the raw data. This may include computer
printouts.
The date is the actual day on which information is entered or
printed on a document.
A written or printed form which is used to furnish information or
Identifiers
Initials
NA or N/A
Overwriting
Performed By
Quarantine
Raw Data
Recorded By
Reviewed By
Signature
Video
Q002
provide instructions.
Information that serves to identify or describe something, such
as effective dates, lot number, line number, equipment number,
manufacturing or task date, product description, container
numbers, specification number and run number. Identifiers can
usually be retrieved from another source or document.
Consist of the first letter of both the first name and the last name,
i.e. surname. Use of the middle initial is optional.
Abbreviation for the phrase “Not Applicable”. It is used to
indicate that the entering of data into a space provided is not
appropriate in that particular case.
Overwriting refers to writing over previously recorded
information to make a change. Overwriting is never allowed on
any cGMP document.
Initials or signature of the person executing an operation or task.
This is usually the “operator” or “analyst”.
The default status for raw materials and packaging components
upon receipt from the supplier and for drug products upon
completion of processing while awaiting evaluation against
identified release criteria.
The actual information obtained from an observation, test,
measurement or activity. This may include computer or
instrument printouts.
Initials or signature of a person documenting information,
results, or readings of an operation. This may be the “operator”.
Initials or signature of the person examining a task, document or
record in order to confirm its accuracy and completeness,
including checking calculations.
Consists of at least the initials of the first name and the complete
last name.
Corporate Quality Concepts: cGMP Documentation Practices
Corporate Quality Concepts: cGMP Documentation Practices
Standard Operating Procedures (SOPs)
SOPs are documents which detail how staff should undertake particular procedures or
processes. These procedures or processes are usually of a general nature, often being
independent of any one pharmaceutical product. Many SOPs fall into one of several
general categories, including:


SOPs detailing step-by-step operational procedures for specific items of
equipment, e.g. autoclaves, homogenizers, freeze-dryers, pH meters, product
labeling machines, etc.;
SOPs detailing maintenance/validation procedures for specific items of equipment
or facility areas, e.g. SOPs detailing CDS (cleaning, decontamination and
sanitation) of clean rooms;


SOPs relating directly to personnel, e.g. step-by-step procedures undertaken when
gowning-up before entering a clean room;
SOPs relating to testing and analysis, e.g. procedures detailing how to sample
properly raw materials or finished products for QC (quality control) analysis,
SOPs relating to the routine sampling and testing of WFI from the ring main
system, etc.
Sections of an SOP:
Purpose
Scope
Responsibilities
References
Definitions
Precautions
Materials/Equipment
Procedure
Attachments
History
General Word of Caution
When Writing SOPs: Keep it general!
Only include what is needed so any qualified person can perform the SOP correctly and
safely. Do not list specific brand names, unless necessary (example: equipment).Give
ranges of times and temperatures if possible. No section should ever be omitted or left
blank. An entry of N/A (not applicable) may be included if there is no information to be
included in a section.
Purpose: Describes why the SOP exists.
Scope: Defines to whom and to what the procedure applies.
Responsibilities: The person or people responsible for performing and updating the SOP.
May also include the person responsible for overseeing the activities of the SOP
References: Other documents which were consulted during the writing of the SOP as
well as those that should be consulted to perform the SOP. Documents may include
manufacturer manuals and other SOPs.
Definitions: Describe any words, phrases or abbreviations which are specific to the SOP.
Commonly used words, phrases or abbreviations need not be described. For example, do
NOT include pH. This is common terminology.
Precautions: Describes any hazards associated with the procedure or with materials used
in performing the procedure.
Materials and Equipment: Any and all materials and/or equipment that are needed to
execute the SOP.
Procedure: A step by step description of the procedure, organized into subgroups.
Attachments: Lists attachments by name and number. Attachments are all documents
that are necessary to perform the SOP. Typically these include diagrams and drawings.
History: Origin of document.
Steps in obtaining an SOP
1.
2.
3.
4.
5.
6.
SOP is written.
Effective date assigned allowing for time to train personnel.
QA (quality assurance) assigns a document number.
Circulated for review.
Approved and signed by QC, QA, operations, and facilities.
QA distributes to authorized personnel. Obsolete versions destroyed. Master copy
retained.
7. Document becomes effective.
LABORATORY EXERCISE NUMBER 1
DOCUMENTATION, QUALITY SYSTEMS AND GMP (PART 1-SOPs)
LABORATORY-WRITING AN SOP
Introduction
Students will study a generic or template SOP and an actual SOP and. These will be used
to write an SOP for use of any piece of equipment with which the students are familiar.
Background
Following is a generic or template SOP. It starts on the next page. On the next page after
the template, and for the next five pages, is an example of an SOP. Study both the
generic/template SOP and the SOP example.
Company Name:
Document Number:
Revision Number:
Issue Date:
Page:
Document Title:
Supersedes: New
Document Originator / Reviser:
Signature
Dept.
__________
Date
I have reviewed this document and find it accurate and complete:
Date:
Date:
This document has been approved as a Master Document.
Date:
Quality Manager
This is an OFFICIAL COPY; a true reproduction of the MASTER
DOCUMENT. It has been checked for accuracy and approved for use.
Date:
Issued by:
1.0
PURPOSE:
1.1
2.0
SCOPE:
2.1
3.0
The scope of this document includes…
RESPONSIBILITIES:
3.1
4.0
The purpose of this document is to describe in detail the…..
It is the responsibility of the Position Title to:


REFERENCES (RELATED DOCUMENTS):
4.1
4.2
Refer to DOC# XX-XXX.
Refer to DOC#XX-XXX.
5.0
DEFINITIONS:
5.1
Definition 1 …
5.2
Definition 2…
6.0
PRECAUTIONS
6.1
Precaution 1……
6.2
Precaution 2……
7.0
MATERIALS AND EQUIPMENT:
7.1
Materials:
7.1.1 Chemical 1….. (Lot #, Product #, handling)
7.1.2 Chemical 2…..
7.2
Equipment:
7.2.1 Hardware 1…
7.2.2 Hardware 2…
.
8.0
PROCEDURE:
8.1
Initial Preparation:
8.1.1 Prepare a ……….
8.1.2 Add the …….
8.1.3 Filter the solution to….
9.0
8.2
Primary Steps:
8.2.1 Measure the…..
8.2.2 Adjust the pH to ……
8.2.3 Centrifuge at ___ g or _____ rpm and…
8.2.4 Resolubilize the …..
8.3
Second Treatment to remove remaining contaminants:
8.3.1 The conductivity is ….
8.3.2 Filtration to …
8.3.3 Pass the filtered solution….
6.3.4 Analyze the pass fraction by SDS-PAGE to determine….
ATTACHMENTS AND EXHIBITS:
9.1
10.0
Attachment or Form #:
HISTORY
Title
Number
A Biotechnology Company
123 Bioscience Drive
Anywhere, US 0007
Document Number: 1.23
Revision Number: 2
Effective Date:
Page 1 of 5
Title: SDS-PAGE SOP
Approvals
Preparer: _________Your Name________________________ Date ________________
Reviewer: ________His Name _______________________ Date ________________
Reviewer: ________His Name _______________________ Date ________________
1. Purpose:
1.1. To describe the appropriate operating instructions to perform SDS-PAGE
analysis of protein samples.
2. Scope:
2.1. Applies to confirming the presence and purity of two recombinant human
proteins (tPA and HSA) produced and purified in the laboratory of A
Biotechnology Company.
3. Responsibilities:
3.1. It is the responsibility of the Supervisor to ensure that this SOP is performed as
described and to update the procedure when necessary.
3.2. It is the responsibility of the technicians to follow the SOP as described and to
inform the Supervisor about any deviations or problems that may occur while
performing the procedure.
4. References:
4.1. Invitrogen Novex Gel instructions
4.2. Novex XCell II Mini-Cell Gel Box Operation SOP
4.3. Gel Documentation Instrument SOP
5. Definitions:
5.1. tPA is tissue plasminogen activator.
5.2. HSA is human serum albumin.
6. Precautions:
6.1. Acrylamide is a neurotoxin. Always wear protective gloves when handling the
polyacrylamide gels.
6.2. Fixative solution is flammable – keep away from sparks and flames. Dispose in
Fixative Hazardous Waste bottle
6.3. GelCode Blue is harmful. Dispose in GelCode Blue Hazardous Waste bottle.
7. Materials:
7.1. Protein Samples
7.2. Protein Standard, 4mg/ml: (2– 8C refrigerator)
7.3. Invitrogen Molecular Weight Marker (2– 8C refrigerator)
7.4. NOVEX Precast Gel Box and Accessories
7.5. Power Supply for Protein Electrophoresis
7.6. NuPAGE 4-12% Bis-Tris Gels (1.0mm x 10 well), 2– 8C refrigerator
7.7. NuPAGE MOPS SDS Running Buffer (20X), room temperature
7.8. NuPAGE Antioxidant, 2– 8C refrigerator
A Biotechnology Company
123 Bioscience Drive
Anywhere, US 0007
Document Number: 1.23
Revision Number: 2
Effective Date:
Page 2 of 5
7.9. NuPAGE SDS Sample Buffer (4X), room temperature
7.10. Reducing Agent (10X), -20C freezer
7.11. Graduated cylinders (1L and 100ml)
7.12. P20, P100 or P200 Micropipettor and tips, including gel loading tips
7.13. Microfuge Tubes
7.14. Microfuge
7.15. Boiling Water Bath
7.16. Staining Trays
7.17. Infors HT Labotron mini rotary shaker
7.18. Gel Fixative Solution
7.19. Pierce GelCode Blue Staining Reagent, 2– 8C refrigerator
7.20. Light Box
7.21. Gel Documentation Instrument
8. Procedure:
8.1. Prepare Running Buffer and Staining Solutions
8.1.1. 1L NuPAGE MOPS SDS Running Buffer (1X) (if needed)
8.1.1.1.Place 50ml NuPAGE MOPS SDS Running Buffer (20X) in a 1 Liter
graduated cylinder.
8.1.1.2. Gently add 950ml deionized water by running it down the side of the
cylinder to make 1 liter of 1X NuPAGE MOPS SDS Running Buffer.
8.1.1.3. Add a stir bar and gently stir. NOTE: SDS is a detergent and will
foam if mixed vigorously. We do not want bubbles.
8.1.2. 200ml NuPAGE MOPS SDS Running Buffer (1X) plus antioxidant (if
needed)
8.1.2.1. Separate 200ml of 1X NuPAGE MOPS SDS Running Buffer into a
500ml Erlenmeyer flask.
8.1.2.2. Add 500µl of NuPAGE Antioxidant.
8.1.2.3. Add a stir bar and gently stir. NOTE: SDS is a detergent and will
foam if mixed vigorously. We do not want bubbles.
8.1.3. 500ml Fixative Solution (if needed)
8.1.3.1. In a 500ml Wheaton bottle, mix together:
250ml 100% Methanol
215ml deionized water
35ml glacial acetic acid
8.1.3.2. Store at 2– 8C
8.2. Dilute Protein Standards (if needed).
8.2.1.1. Dilute the appropriate protein standard(s) with water to a final
concentration of 1mg/ml
8.2.1.2. Label tube with protein name, 1mg/ml, [date], [initials]. Store on ice
until ready to use.
A Biotechnology Company
123 Bioscience Drive
Anywhere, US 0007
Document Number: 1.23
Revision Number: 2
Effective Date:
Page 3 of 5
8.3. Prepare Protein Samples and Protein Standards
(Do NOT perform this step with the Molecular Weight Marker)
8.3.1. For all the samples and the standards, combine the following in a sterile
1.5ml microfuge tube: 25ul 4x sample buffer
10ul 10x reducing agent
65ul sample
8.3.2. Mix gently with a pipette by aspirating and dispensing at least 3 times
8.3.3. Boil for 3-5 minutes.
8.3.4. Remove from boiling water bath.
8.3.5. Pulse all samples and standards in a microfuge for 30 seconds.
8.4. Prepare Novex Precast Gel Box
8.4.1. Assemble gel box according to its SOP.
8.4.2. Place 200ml NuPAGE MOPS SDS Running Buffer (1X) plus antioxidant
in the upper buffer chamber (small chamber between 2 gels or the gel and
buffer dam)
8.4.3. Fill the lower buffer chamber with approximately 600ml of 1X NuPAGE
MOPS-SDS Running Buffer (large chamber).
8.4.4. Rinse gel wells with micropipettor and buffer from upper buffer chamber.
8.5. Load Samples
8.5.1. Using a micropipettor and disposable tips, load 10ul of the Molecular
Weight Marker into one well and up to 50µl of each sample into separate
wells.
8.5.2. Load any empty wells with 15µl of diluted 4X Sample Buffer.
8.5.3. Record order of samples and volumes loaded.
8.6. Run NOVEX NuPAGE MOPS SDS Precast Gel Box
8.6.1. Plug electrophoresis chamber into the gel electrophoresis power supply.
8.6.2. Run gel at 200V for 40 – 60 minutes.
8.6.3. Turn off the power supply when the dye reaches 1cm from the bottom of
the gel.
8.7. Stain and Photodocument the NOVEX NuPAGE MOPS SDS Precast Gel
8.7.1. Disassemble gel box per SOP and remove gel from plastic cassette.
8.7.2. Rinse gel box well with DI water. Do not use brushes on the gel box, they
scratch the surface. Do not immerse top of gel box or electrical
components.
8.7.3. Place gel in staining tray.
8.7.4. Wash gel 3 times for approximately 5 minutes with DI water at room
temperature on a shaker.
8.7.5. Add enough Fixative solution to cover the gel completely and fix for
approximately 15 minutes at room temperature on a shaker.
8.7.6. Discard Fixative Solution into the Fixative Hazardous Waste bottle.
A Biotechnology Company
123 Bioscience Drive
Anywhere, US 0007
Document Number: 1.23
Revision Number: 2
Effective Date:
Page 4 of 5
8.7.7. Wash gel 3 times for a minimum of 5 minutes with DI water at room
temperature on a shaker.
8.7.8. Add about 50ml of GelCode Blue and stain for 1-24 hours at room
temperature on a shaker.
8.7.9. Decant GelCode Blue into GelCode Blue Hazardous Waste bottle.
8.7.10. Wash gel with DI water for 15 minutes to overnight on a shaker
8.7.11. Remove gel from staining tray and place on visible light box
8.7.12. Identify the protein standards and samples and estimate their molecular
weights. See Molecular Weight Diagram.
9. Attachments:
9.1. Molecular Weight Marker Diagram
10. History:
Name
John Smith
A Person
Her Name
Date
2002
2005
2007
Amendment
Initial Release
Changed Coomassie stain to GelCode Blue Stain
Put into SOP 2005 format
A Biotechnology Company
123 Bioscience Drive
Anywhere, US 0007
Attachment: Molecular Weight Maker Diagram
Document Number: 1.23
Revision Number: 2
Effective Date:
Page 5 of 5
SOP Exercise
Select any item of laboratory equipment the use of which you are familiar with. It need
not be anything complicated. For example a pH meter, a conductivity meter, a balance or
even a hot plate and stirrer would be suitable pieces of equipment. Using the above
generic or template SOP and the example SOP as guides, write an SOP for operation of
the item of laboratory equipment that you have chosen.
Once you have obtained all the information you need from the laboratory, this exercise
may be completed outside of the laboratory. Write up and print the SOP as if it was the
genuine document. Use the format in the generic/template SOP for the preparer and
reviewers’ signatures and acceptance of the SOP. This would make the document the best
possible and would indicate approval by two reviewers, acceptance by the Quality
Manager, and final issuance by the Company. All these features are lacking in the
example SOP. Ask classmates to be the two reviewers, the Quality Manager and the
issuer.
LABORATORY EXERCISE NUMBER 2
DOCUMENTATION, QUALITY SYSTEMS AND GMP (PART 2-BATCH
RECORD AUDIT)
Introduction
Students will gain an understanding of good documentation and cGMPs by the audit of a
batch record.
Background
Each of the Master Batch Production Record (MBPR) and the Batch Production
Record (BPR) is a DETAILED step-by-step of the entire production process for a batch
of product. It includes:


Types and quantities of components and raw materials.
Processing parameters such as
o Incubation times
o Run times
o When to add components and in what order, i.e. complete instructions on
adding, mixing and sampling for Quality Control Unit (QC) testing
o Expected yields



Processing quality control parameters such as what QC tests to perform.
Environmental control parameters such as the type of clean room required.
Specifications for packaging of the product and label(s) that will be used.
The Master Batch Production Record is kept and provides the individual Batch
Production Records.
The Batch Production Record:
 Accurately follows the Master Batch Production Record.
 Includes Quality Control Unit review and approval of
o Batch Production Record
o Cross reference of receiving and Batch Production Records
o Any material review and disposition decision
o Reprocessing
o Release for distribution


Is kept for 3 years beyond the date of batch production.
Includes, in part,
o Batch, lot or control number of product
o Identity of equipment and processing lines used
o Date and time of the maintenance, cleaning and sanitizing of the equipment
and processing lines used
o
o
o
o
o
o
o
o
Incoming shipment lot identifier
Identity and mass or measure of each component used
Date and initials of persons completing and verifying steps.
Date batch produced.
Test results.
Any material reviews and disposition decisions
Documentation that final product specifications are met.
Copy(ies) of label(s) used for the packaged product.
The batch production record, or more simply batch record, accompanies a product as it is
made. The batch record directs the operators in exactly how to make the product – and
the operators must follow the batch record instructions just as it is written. Each time a
product is to be made, the operators are issued a fresh copy of the current version of the
batch record. The batch record also provides blanks that are filled in as the operator
performs each task to document that they have done it. For critical steps, a witness
watches the operator and signs off as well. By filling in the blanks properly, the operators
demonstrate that they have done each task properly. Other points of note for batch
records are as follows (some have already been mentioned above).
COMPANY NAME: The name of the company should be included on the batch record.
TITLE: The title of the batch record is included on each page.
ID NUMBER: Each batch record should also contain a unique identification number. It
may be listed as the “Batch Record Number.”
PAGE NUMBER: Each page of the batch record should be numbered.
HAZARD COMMUNICATION: This section warns the operator of any hazards
associated with the procedure and any required safety precautions.
PROCEDURE: The core of the batch record details what the operator will do in a stepby-step chronological manner. Every batch record has this information. As the operator
performs each task, s/he fills in related information to document how that task was done
and initials the “operator” blank. The witness initials the “verified by” blank. The date
must also be provided for the initials in each case, i.e. for both “operator” and “verified
by.”
LABEL INFORMATION: Every company will have specific instructions for labeling
manufactured products. These instructions must be followed exactly to avoid any mixups.
QA REVIEW: Every completed batch record is reviewed by a QA (Quality Assurance)
representative to ensure that it is properly filled out.
Laboratory Activity
On the following page is the front page of a batch record drawn up by a hypothetical
company producing small quantities of E. coli media as one of its activities. As part of
the company’s quality system you are required to audit this batch record before it is
approved and issued to the technicians. Based on the above discussion of batch records,
there are at least five omissions in the one below. Draw up and submit a list of these.
LABORATORY EXERCISE NUMBER 3
DOCUMENTATION, QUALITY SYSTEMS AND GMP (PART 3-BATCH
RECORDS)
Introduction
Students will practice good documentation and cGMP in two laboratory activities
(Laboratory Exercise Numbers 3 and 4) designed as an introduction to manufacturing.
Materials and Supplies (To Be Used in Laboratory Exercise Number 4)
Bread; peanut butter; jelly; utensils (knives, spoons); aluminum foil; balances
Activity
Work in groups to develop a company that will manufacture peanut butter and jelly
sandwiches. Each group will develop the following in this first activity:
Part A
1. Name of Firm.
2. Quality Policy and Mission Statement.
3. Organizational Chart.
Part B
1. Specifications for Sandwiches.
2. Raw Materials Needed For Sandwiches.
3. Equipment Requirements.
4. Process Controls.
For Part A, use any resources, such as the websites of biotechnology companies
manufacturing products, that you believe would be of assistance.
Retain all you have developed from above for the second activity, in Laboratory Exercise
Number 4.
LABORATORY EXERCISE NUMBER 4
DOCUMENTATION, QUALITY SYSTEMS AND GMP (PART 4-BATCH
RECORDS, CONTINUED)
Introduction
This is a continuation from Laboratory Exercise Number 3.
Materials and Supplies
Bread; peanut butter; jelly; utensils (knives, spoons); aluminum foil; balances
Activity
Part A
1. Review batch records (batch production records) in Laboratory Exercise Number 2.
2. Each company creates a batch record for the manufacture of its product, viz. peanut
butter and jelly sandwiches.
Part B
1. Each group gives its batch record to another company for production of its product.
2. Each group manufactures the product and evaluates the batch record during the
process.
Part C
1. Each group reviews the quality of its product with the group which performed the
manufacturing.
2. Discuss the quality of the batch record and any modifications thereof.
Report
Each group will submit a report containing all the information required in Parts A and B
of Laboratory Exercise Number 3. In addition, each group will include in its report batch
records from above; one prior to and one after modifications with a discussion of the
quality of the batch record in each case.
LABORATORY EXERCISE NUMBER 5
GOWNING
Introduction
In a biomanufacturing facility, gowning requirements for operational areas are set
appropriately for the activities that occur within those areas. Appropriate gowning will
minimize the number of particulates shed by each operator. This helps a facility maintain
the appropriate air classification in each area, which in turn protects the product from
potential contamination.
Gowning
Gowning is the donning of clean room garments to protect the low particle counts
required in the processing areas. Different levels of gowning are required, depending on
the operations being performed.
Code of Federal Regulations
21CFR 211.28(a) states: Personnel engaged in the manufacture, processing, packing, or
holding of a drug product shall wear clean clothing appropriate for the duties they
perform. Protective apparel, such as head, face, hand, and arm coverings, shall be worn
as necessary to protect drug products from contamination.
In General
General gowning points include the following:
 Gowning will take place in gowning air locks.
 Each gowning room will have the maximum number of individuals allowed to
gown at one time clearly posted.
 Choose a gown that is large enough to allow unrestricted movement, but not so
large as to interfere with movement.
Disposable or Reusable Gowns?
In some areas, disposable gowns will be used instead of reusable garments. Disposable
gowns should be placed in trash receptacles upon degowning. Reusable garments should
be placed in laundry bins upon degowning.
Permitted Items
The following items are permitted in gowning level 0-3 areas:
 Earrings
 Rings
 Watches
Prohibited Items
The following items are not permitted in the manufacturing facility at any time:
 Cosmetics that could shed particles
 Excessive jewelry that cannot be covered by gowning, for example dangling
earrings.
Key Concepts
The following points are key concepts:
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In the locker rooms, prior to entering the circulation corridor, shoes should be
inspected for mud and debris. If necessary, replace soiled shoes with clean shoes.
Tacky mats are usually present near gowning room doors. Whenever moving
through a gowning room, always step on the tacky mat to remove debris from the
bottoms of your shoes.
Prior to donning appropriate level gowning, hands must be sanitized. Hand
sanitization stations are available in each gowning room.
Safety glasses must be donned prior to entering all processing rooms within the
production areas. Safety glasses are required and can be sanitized with ethanol
prior to wearing. Safety glasses are not required in non-processing rooms within
the production areas.
If any gowning garment is found to be defective, it should be tied in a knot for
easy identification and placed in the soiled garment bin.
While gowning, every effort should be made to minimize garment contact with
floor, wall or equipment.
When gowning, all zippers and snaps should be completely fastened. If they are
not, the gown will not perform as it is designed.
At various points in or around the gowning area there are lines of demarcation on
the floor. These demarcation lines separate the clean side from the dirty side of
the gowning room. Shoe covers and knee-high boots are donned as you step
across the line of demarcation from the dirty side to the clean side. If one has
stepped back into the dirty side with the intention of returning to the clean side,
the shoe covers or boots must be removed, discarded in the soiled garment bin,
and replaced with new covers or boots on stepping back into the clean side.
To degown, reverse the appropriate level gowning sequence. For areas in which
single-use garments are worn, operators may step over the line of demarcation to
a lower gown level to remove gown garments.
Only one activity, e.g. gowning, degowning, material transfer, environmental
monitoring, cleaning, etc., may occur in a gown or degown room at a time.
For mechanical space that is accessed from GMP process areas, personnel may be
required to don additional gown garments, as posted, and remove those garments
upon reentry into a process area.
GOWNING LEVELS 0-3
Introduction
Appropriate gowning requirements are determined for any given biomanufacturing
facility. Gowning levels are presented as an example of gowning for a manufacturing
facility. However, they are not a universal industry standard.
Gowning Levels 0 Through 3
The following are the minimum requirements for gowning Levels 0 through 3:
 Level 0 gowning consists of street clothes (clean pants, shirts with sleeves, and
clean, close-toed shoes) or plant uniforms and safety glasses.
 Level 1 gowning consists of items in gowning level 0 plus shoe covers.
 Level 2 gowning consists of items in gowning level 1 plus a frock, hair cover and
facial hair cover, when appropriate.
 Level 3 gowning consists of items in gowning level 0 plus knee-high boots, hair
cover, facial hair cover, when appropriate, and a coverall.
Level 0
Gown for Level 0 in the following sequence:
Action
Inspect shoes for mud and debris. Replace
with clean shoes if necessary.
Sanitize safety glasses with a disinfectant,
such as 70% isopropyl alcohol, or prewetted wipes.
Ensure that legs and shoulders are covered
and that facility scrubs are available, if
necessary.
Rationale
Depositing debris in the circulation
corridor increases the chance of
contaminating the processing areas.
Removes bacteria and debris that may be
carried into the manufacturing areas.
This is necessary to minimize particulate
shedding within the facility.
Level 0 Applicability
Refer to the applicable facility gowning Standard Operating Procedure for Level 0
gowning applicability.
Level 1
Gown for Level 1 in the following sequence:
Action
Sanitize hands with Alcare or equivalent.
Sanitize safety glasses with a disinfectant,
such as 70% isopropyl alcohol, or prewetted wipes.
Don shoe covers while stepping across the
line of demarcation from the dirty to the
clean side of the gowning room.
Rationale
Clean gowning should be handled with
clean hands.
Removes bacteria and debris that may be
carried into the manufacturing areas.
This action ensures that the clean side of
the gowning room remains clean.
Level 1 Applicability
Refer to the applicable facility gowning Standard Operating Procedure for Level 1
gowning applicability.
Level 2
Gown for Level 2 in the following sequence:
Action
Sanitize hands with Alcare or equivalent.
Sanitize safety glasses with a disinfectant,
such as 70% isopropyl alcohol, or prewetted wipes.
Don hair cover and tuck in all hair.
Don facial hair cover, if applicable.
Don shoe covers while stepping across the
line of demarcation from the dirty to the
clean side of the gowning room.
Don appropriately-sized frock and ensure
that all zippers and snaps are completely
closed.
Rationale
Clean gowning should be handled with
clean hands.
Removes bacteria and debris that may be
carried into the manufacturing areas.
Hair covers keep hair from shedding.
Hair covers keep hair from shedding.
This action ensures that the clean side of
the gowning room remains clean.
Gown will not do its job properly if it is not
completely closed.
Level 2 Applicability
Refer to the applicable facility gowning Standard Operating Procedure for Level 2
gowning applicability.
Level 3
Gown for Level 3 in the following sequence:
Action
Sanitize hands with Alcare or equivalent.
Sanitize safety glasses with a disinfectant,
such as 70% isopropyl alcohol, or prewetted wipes.
Don hair cover and tuck in all hair.
Don facial hair cover, if applicable.
Don knee-high boots while stepping across
the line of demarcation from the dirty to the
clean side of the gowning room.
Don appropriately-sized coverall and tuck
legs into knee-high boots.
Rationale
Clean gowning should be handled with
clean hands.
Removes bacteria and debris that may be
carried into the manufacturing areas.
Hair covers keep hair from shedding.
Hair covers keep hair from shedding.
This action ensures that the clean side of
the gowning room remains clean. Donning
boots first keeps the inside of the coverall
clean.
Tucking ensures that gown legs will not
touch the floor and become soiled and
keeps particulates from escaping down the
gown leg on to the floors.
Level 3 Applicability
Refer to the applicable facility gowning Standard Operating Procedure for Level 3
gowning applicability. Level 3 gowning is required in areas where processing occurs,
with the addition of a mask and gloves when appropriate. It is considered good practice to
wear gloves at all times within the processing areas, but gloves are not required for
minimum Level 3 gowning. Gloves, however, must be worn whenever equipment is
handled within the production areas. If at any time gowning becomes visibly soiled,
personnel must move to a degowning room, remove the soiled gown, and reenter the area
through the gowning room, donning appropriate clean gowning material. Once inside the
processing area, personnel may not unzip their coverall or degown in any way, except in
case of emergency. Personnel must move to a gowning room to access cell phones,
radios, etc. within the gown.
Masks and Gloves
When performing certain operations, a mask and/or gloves must be worn in addition to
Level 3 gowning:
When
Handling clean or sanitized labware or
product-processing components.
Product or processing materials are
exposed to the environment.
Taking a sample of in-process product.
Making aseptic product contact
connections.
Then
Wear clean gloves.
All individuals in the clean room must
wear masks and clean gloves.
Individual taking the sample must wear a
mask and clean gloves.
Individual making the connections must
wear a mask and clean gloves.
GOWNING
LABORATORY
Introduction
Students will learn the concepts and skills involved in gowning for both outside and
inside an aseptic area (Class 100). Each student will don complete gowning attire in an
aseptic manner.
Materials and Supplies
Hand-sanitizing agent (e.g. Alcare)
70% isopropyl alcohol
Wipes
Safety glasses
Gowns: Frocks and Coveralls
Boots
Hoods
Gloves
Masks
Beard covers
Shoe covers
Bouffant caps
Activities
1. The laboratory activities will begin with a demonstration of gowning and gloving
techniques by the industry instructor. Gowning for both outside and inside the aseptic
area of a clean room will be demonstrated.
2. The sequences for degowning will also be demonstrated and performed.
3. Students will perform the gowning and degowning sequences under supervision.
4. Students may critique the performance of each other.
LABORATORY EXERCISE NUMBER 6
AUTOCLAVE MONITORING
Introduction
Autoclaves are large pieces of equipment that are widely used in the biotechnology
industry. They contain large compartments capable of sterilizing a wide variety of
equipment and labware used in the production process. While disinfectants are used in
the sanitization of floors, walls, tabletops and other surfaces, the autoclave is the means
by which labware and other pieces of equipment are sterilized for use in applications
where sterility is required. The autoclave is capable of sterilizing many pieces of
equipment in a relatively short period of time. Equipment and labware are cleaned prior
to autoclaving and the autoclave is used to sterilize the equipment. At 121.1°C for a
specified length of time, usually 15 to 20 minutes depending on the equipment,
microorganisms and their spores are no longer viable. The autoclave, through the
injection of clean steam, raises the pressure and temperature of its interior, and therefore
its contents, to at least this level. Equipment or labware can be considered sterile and
ready for use once it has been sustained at this temperature for the required period of
time.
Equipment Validation
For use in a GMP environment, equipment such as an autoclave would need to be
validated. Equipment Validation describes the inspection and qualification of GMP
equipment and the associated documentation to verify that predetermined fabrication,
installation and operational specifications are met. Equipment Validation ensures that an
instrument is appropriate for its intended use. Examples of equipment that must be
validated are:
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Steam autoclaves
Dry heat ovens
Depyrogenation tunnels
Ethylene oxide (ETO) sterilizers
Freeze dryers
Incubators
Refrigerators
Pumps
HEPA filters
Chromatography
Fermentors
Typical validation phases are:
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Design qualification (DQ) for setting functional and performance specifications
(operational specifications).
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Installation qualification (IQ) for performing and documenting the installation in
the selected user environment.
Operational qualification (OQ) for testing the equipment in the selected user
environment to ensure that it meets the previously defined functional and
performance specifications.
Performance qualifications (PQ) for testing that the system consistently performs
as intended for the selected application.
The validation of a GMP manufacturing facility is a complex operation. Many associated
documents require pre- and post-validation approval. Qualified staff must perform the
inspections and performance tests in the facility. Good planning, resource allocation and
test design are required for the successful deployment of validation activities and the
timely completion of reports.
Once the initial validation is concluded, the equipment must be monitored to ensure that
it remains in a validated state throughout its use, i.e. equipment must be periodically
tested to ensure that it is still operating within its pre-determined specifications.
Revalidation must be performed after modifications to ensure the same.
The steps of validation are:
1. System Documentation
2. SOPs (operation, calibration, maintenance, sampling and testing)
3. Write Documentation (IQ/OQ/PQ) with input from:
 Engineering
 Facilities
 Manufacturing
 Quality Assurance (QA)
 Regulatory Affairs (RA) (often as policies)
4. Execution of IQ/OQ/PQ in Facility
5. Review and Approvals of the Validation Technical Reports (by same functions that
approved the Validation Protocols)
To get validation done:
 A Validation Protocol needs to be developed.
 The validation procedures according to the protocols (IQ, OQ, PQ) need to be
performed and documented.
 The Validation Technical Report needs to be signed and issued.
A Validation Protocol is an experimental plan intended to produce documented evidence
that a system has been validated. Types of Validation Protocols:
1. Installation Qualification (IQ). An IQ is documented verification that all key aspects
of the installation adhere to appropriate codes and approved designs and conform to
the production department’s and the manufacturer’s specifications.
2. Operational Qualification (OQ). An OQ is documented verification that the
equipment performs as intended throughout all anticipated operating ranges.
3. Process Qualification (PQ). A PQ is documented evidence which provides a high
degree of assurance that a specific process will consistently produce a product that
meets its predetermined specifications and quality attributes.
In executing the Validation Protocols:
 The procedures described in the respective SOP need to be followed carefully and
documented fully.
 The validation work continues until the acceptance criteria are met.
 If acceptance criteria cannot be met, the suitability of the equipment or of the
specified criteria needs to be discussed.
In the Technical-Summary Report from the Validation Protocol:
 The validation study’s goals and approach should be summarized.
 The results should be summarized.
 Deviations from the original acceptance criteria should be explained and justified.
 Approval signatures are required.
The monitoring of an autoclave in the present laboratory exercise does not in itself
constitute an Operational Qualification (OQ), which would be the pertinent Validation
Protocol documenting that that the autoclave operates according to its predetermined
specifications. However, this monitoring could provide some of the information
necessary for an OQ. Other information would be required such as:
 SOP and trainer verification
 Operators’ manual
 Start-up and shut down
 Alarms test
 Instrumentation calibration
 Integrity of vessel and seals
 Emergency recovery, after loss of power and/or services
Objectives
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To monitor the saturated steam sterilization cycle at 121°C of an autoclave using
a biological indicator.
To gain familiarity with a task, i.e. the evaluation of sterilization by an autoclave,
that could be used for validation of the operation of the autoclave, i.e. OQ.
Materials
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Autoclave.
Raven ProSpore Self-Contained Biological Indicator. Each ampoule contains a
suspension of spores of the organism, Geobacillus stearothermophilus, within a
growth medium also containing Bromocresol Purple to function as a pH indicator.
The acid production associated with growth causes a change in color from purple
to or toward yellow.
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Incubator set at 55-60°C.
Procedure
1. Exposure. Add about 50 ml water to a 250 ml beaker. Suspend an ampoule of the
Raven ProSpore Biological Indicator in the water contained in the beaker. This is
done because sterilization of an item would be less effective if it is in water. If it is
functioning correctly, the autoclave would adequately sterilize water and anything
contained within it. If the autoclave was not functioning properly it would be detected
most markedly with the biological indicator autoclaved in water. Place the beaker of
water containing the biological indicator near the drain of the autoclave. This is
usually the most difficult location to sterilize and the biological indicator is placed
here for the same reason it is placed in water. Run the autoclave cycle.
2. Caution. On completion of the autoclave cycle unlock the autoclave door and open it
carefully. Remove the beaker containing the ampoule. Contents of the ampoule are
hot and under pressure and therefore it must be handled with care. Allow a sufficient
cooling time of 10-15 minutes. Failure to do so may result in bursting of the ampoule.
3. Incubation. Place the processed ampoule in a vertical position in an incubator at 5560°C. Mark a control ampoule, i.e. one that was not autoclaved, and incubate along
with the processed ampoule to ensure spore viability. Incubate for 48 hours.
4. Monitoring. Examine the Prospore ampoules daily during incubation. Record your
observations. All positive ampoules should be recorded and then disposed of
immediately.
5. Interpretation.
Control: The control ampoule should exhibit a color change to or towards yellow
and/or turbidity. If the control ampoule does not show signs of growth consider
the test invalid.
Test: A failed sterilization cycle is indicated by turbidity and/or a change in color
to or towards yellow. A test ampoule that retains its purple color and is not turbid
indicates an adequate sterilization cycle.
6. Results. Record all your observations and state your conclusions in your laboratory
report.
LABORATORY EXERCISE NUMBER 7
GMP POPCORN
Goal
Students will make, with adherence to GMP (Good Manufacturing Practices), a batch of
popcorn. This must be completed within a fixed time of 2 hours.
Objectives
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To achieve further understanding of the complexity of a GMP process.
Simulation of a biomanufacturing facility and the different departments that
would interact therein.
To be able to complete all the necessary GMP documentation.
An appreciation of how working in teams and cooperation between departments
are essential features of GMP.
To gain insight into the pressures and constraints that one could face while
working in industry.
This Exercise Could Reflect Industry
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Frustration could be experienced at times.
There may be a feeling of being rushed.
There may be a sense of accomplishment once the batch has been made.
The actual time for the process, which in industry would be such things as
extraction, fermentation, purification, etc., is very small compared to the time it
takes to get all GMP documentation in place.
Rewards for completion of the task will be in the form of a salary increase and/or
stock options.
Materials
1. Paper for documentation
2. Approved labels
3. Quarantine labels
4. Box for approved and quarantine
5. Microwave Popcorn
6. Access to microwave oven
7. Water in a squirt bottle
8. Cleaning agent
9. Paper towels
10. Sponge
11. Measuring cups
12. Bags for the finished product
Teams
A)
B)
C)
D)
Material Control: 2 people
Quality Control (QC): 2 people
Quality Assurance (QA): 2 people
Production: 4 people
Students should each join a particular team. If numbers are sufficient, distribute
yourselves according to the numbers given above. Otherwise ensure that there is at least
one person in each team except for production where at least two people are preferred.
Documentation
Inform the instructor when you have completed production and the product is ready to be
shipped. A bonus of $10,000.00 in company stock options (equivalent to 10% on the final
grade for this laboratory exercise) for all the members of all the teams will be awarded
for every 15 minutes that the product is obtained ahead of schedule with all the
documentation completed, i.e. before the end of the 2 hour time limit. A further bonus of
a 10% salary increase (equivalent to 10% on the final grade) will be awarded if the
product is of good quality.
Submit all documentation together with the names of the individuals assigned to each
named team.