Document Sample



1                     DOCUMENTATION, QUALITY
                      SYSTEMS AND GMP (PART 1-SOP

2                     DOCUMENTATION, QUALITY
                      SYSTEMS AND GMP (PART 2-BATCH
                      RECORD AUDIT)

3                     DOCUMENTATION, QUALITY
                      SYSTEMS AND GMP (PART 3-BATCH

4                     DOCUMENTATION, QUALITY
                      SYSTEMS AND GMP (PART 4-BATCH
                      RECORDS, CONTINUED)

5                     GOWNING

6                     AUTOCLAVE MONITORING

7                     GMP POPCORN


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

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

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

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

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.


   Approved By        Signature of a qualified individual (supervisor or designee)
                      indicating that the information documented is complete,
                      accurate, and acceptable.
   Backdating         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.
   Batch              Collection of records associated with the manufacture of a
   Production         specific lot of product.
   Comment            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.
   Controlled         Written approved documents used in association with cGMP-
   Documents          related activities to ensure compliance with US and international
                      regulations, as well as company standards.
   Cross-out          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.
   Data               The values and information generated by processing, calculating
                      or transcribing from the raw data. This may include computer
   Date               The date is the actual day on which information is entered or
                      printed on a document.
   Document           A written or printed form which is used to furnish information or
                     provide instructions.
  Identifiers        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.
  Initials           Consist of the first letter of both the first name and the last name,
                     i.e. surname. Use of the middle initial is optional.
  NA or N/A          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        Overwriting refers to writing over previously recorded
                     information to make a change. Overwriting is never allowed on
                     any cGMP document.
  Performed By       Initials or signature of the person executing an operation or task.
                     This is usually the “operator” or “analyst”.
  Quarantine         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.
  Raw Data           The actual information obtained from an observation, test,
                     measurement or activity. This may include computer or
                     instrument printouts.
  Recorded By        Initials or signature of a person documenting information,
                     results, or readings of an operation. This may be the “operator”.
  Reviewed By        Initials or signature of the person examining a task, document or
                     record in order to confirm its accuracy and completeness,
                     including checking calculations.
  Signature          Consists of at least the initials of the first name and the complete
                     last name.
  Video              Corporate Quality Concepts: cGMP Documentation Practices
  Q002               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:


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. SOP is written.
2. Effective date assigned allowing for time to train personnel.
3. QA (quality assurance) assigns a document number.
4. Circulated for review.
5. Approved and signed by QC, QA, operations, and facilities.
6. QA distributes to authorized personnel. Obsolete versions destroyed. Master copy
7. Document becomes effective.





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.


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:

Document Title:
Supersedes: New

           Document Originator / Reviser:

                  Signature                         Dept.                Date

           I have reviewed this document and find it accurate and complete:



           This document has been approved as a Master Document.

                  Quality Manager

           This is an OFFICIAL COPY; a true reproduction of the MASTER
           DOCUMENT. It has been checked for accuracy and approved for use.

                  Issued by:
1.0   PURPOSE:

      1.1   The purpose of this document is to describe in detail the…..

2.0   SCOPE:

      2.1   The scope of this document includes…


      3.1   It is the responsibility of the Position Title to:


      4.1   Refer to DOC# XX-XXX.
      4.2   Refer to DOC#XX-XXX.

      5.1  Definition 1 …
      5.2  Definition 2…

      6.1 Precaution 1……
      6.2 Precaution 2……


      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…


      8.1   Initial Preparation:
            8.1.1 Prepare a ……….
            8.1.2 Add the …….
            8.1.3 Filter the solution to….
       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….


       9.1   Attachment or Form #:     Title

10.0   HISTORY
A Biotechnology Company                                      Document Number: 1.23
123 Bioscience Drive                                         Revision Number: 2
Anywhere, US 0007                                            Effective Date:
                                                             Page 1 of 5

                             Title: SDS-PAGE SOP
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                                      Document Number: 1.23
123 Bioscience Drive                                         Revision Number: 2
Anywhere, US 0007                                            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)
  50ml NuPAGE MOPS SDS Running Buffer (20X) in a 1 Liter
                   graduated cylinder.
  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.
  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
  Separate 200ml of 1X NuPAGE MOPS SDS Running Buffer into a
                   500ml Erlenmeyer flask.
  Add 500µl of NuPAGE Antioxidant.
  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)
  In a 500ml Wheaton bottle, mix together:
                       250ml 100% Methanol
                       215ml deionized water
                       35ml glacial acetic acid
  Store at 2– 8C
   8.2. Dilute Protein Standards (if needed).
  Dilute the appropriate protein standard(s) with water to a final
                   concentration of 1mg/ml
  Label tube with protein name, 1mg/ml, [date], [initials]. Store on ice
                   until ready to use.
A Biotechnology Company                                   Document Number: 1.23
123 Bioscience Drive                                      Revision Number: 2
Anywhere, US 0007                                         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
       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
       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                                    Document Number: 1.23
123 Bioscience Drive                                       Revision Number: 2
Anywhere, US 0007                                          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            Date       Amendment
John Smith      2002       Initial Release
A Person        2005       Changed Coomassie stain to GelCode Blue Stain
Her Name        2007       Put into SOP 2005 format
A Biotechnology Company                      Document Number: 1.23
123 Bioscience Drive                         Revision Number: 2
Anywhere, US 0007                            Effective Date:
                                             Page 5 of 5

Attachment: Molecular Weight Maker Diagram
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



Students will gain an understanding of good documentation and cGMPs by the audit of a
batch record.


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   Incoming shipment lot identifier
       o   Identity and mass or measure of each component used
       o   Date and initials of persons completing and verifying steps.
       o   Date batch produced.
       o   Test results.
       o   Any material reviews and disposition decisions
       o   Documentation that final product specifications are met.
       o   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 step-
by-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
LABEL INFORMATION: Every company will have specific instructions for labeling
manufactured products. These instructions must be followed exactly to avoid any mix-
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.



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


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.



This is a continuation from Laboratory Exercise Number 3.

Materials and Supplies

Bread; peanut butter; jelly; utensils (knives, spoons); aluminum foil; balances


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

Part C
1. Each group reviews the quality of its product with the group which performed the
2. Discuss the quality of the batch record and any modifications thereof.


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.



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 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

Key Concepts

The following points are key concepts:

      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


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                                        Rationale
Inspect shoes for mud and debris. Replace      Depositing debris in the circulation
with clean shoes if necessary.                 corridor increases the chance of
                                               contaminating the processing areas.
Sanitize safety glasses with a disinfectant,   Removes bacteria and debris that may be
such as 70% isopropyl alcohol, or pre-         carried into the manufacturing areas.
wetted wipes.
Ensure that legs and shoulders are covered     This is necessary to minimize particulate
and that facility scrubs are available, if     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                                         Rationale
Sanitize hands with Alcare or equivalent.      Clean gowning should be handled with
                                               clean hands.
Sanitize safety glasses with a disinfectant,   Removes bacteria and debris that may be
such as 70% isopropyl alcohol, or pre-         carried into the manufacturing areas.
wetted wipes.
Don shoe covers while stepping across the      This action ensures that the clean side of
line of demarcation from the dirty to the      the gowning room remains clean.
clean side of the gowning room.

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                                         Rationale
Sanitize hands with Alcare or equivalent.      Clean gowning should be handled with
                                               clean hands.
Sanitize safety glasses with a disinfectant,   Removes bacteria and debris that may be
such as 70% isopropyl alcohol, or pre-         carried into the manufacturing areas.
wetted wipes.
Don hair cover and tuck in all hair.           Hair covers keep hair from shedding.
Don facial hair cover, if applicable.          Hair covers keep hair from shedding.
Don shoe covers while stepping across the      This action ensures that the clean side of
line of demarcation from the dirty to the      the gowning room remains clean.
clean side of the gowning room.
Don appropriately-sized frock and ensure       Gown will not do its job properly if it is not
that all zippers and snaps are completely      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                                          Rationale
Sanitize hands with Alcare or equivalent.       Clean gowning should be handled with
                                                clean hands.
Sanitize safety glasses with a disinfectant,    Removes bacteria and debris that may be
such as 70% isopropyl alcohol, or pre-          carried into the manufacturing areas.
wetted wipes.
Don hair cover and tuck in all hair.            Hair covers keep hair from shedding.
Don facial hair cover, if applicable.           Hair covers keep hair from shedding.
Don knee-high boots while stepping across       This action ensures that the clean side of
the line of demarcation from the dirty to the   the gowning room remains clean. Donning
clean side of the gowning room.                 boots first keeps the inside of the coverall
Don appropriately-sized coverall and tuck       Tucking ensures that gown legs will not
legs into knee-high boots.                      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                                               Then
Handling clean or sanitized labware or          Wear clean gloves.
product-processing components.
Product or processing materials are             All individuals in the clean room must
exposed to the environment.                     wear masks and clean gloves.
Taking a sample of in-process product.          Individual taking the sample must wear a
                                                mask and clean gloves.
Making aseptic product contact                  Individual making the connections must
connections.                                    wear a mask and clean gloves.



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
Safety glasses
Gowns: Frocks and Coveralls
Beard covers
Shoe covers
Bouffant caps


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.



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

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:

      Steam autoclaves
      Dry heat ovens
      Depyrogenation tunnels
      Ethylene oxide (ETO) sterilizers
      Freeze dryers
      Incubators
      Refrigerators
      Pumps
      HEPA filters
      Chromatography
      Fermentors

Typical validation phases are:

      Design qualification (DQ) for setting functional and performance specifications
       (operational specifications).
      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


      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.


      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.
      Incubator set at 55-60°C.


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 55-
   60°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
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

                      GMP POPCORN


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.


      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

      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.


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

   A)   Material Control: 2 people
   B)   Quality Control (QC): 2 people
   C)   Quality Assurance (QA): 2 people
   D)   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.


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.

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