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VALIDATION of FREEZING BAGS AN SOP I am not independent, I manufacture freezing bags I am President of American Fluoroseal Corporation (Afc). – PowerPoint PPT presentation

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  • AN SOP

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I am not independent, I
manufacture freezing bags
  • I am President of American Fluoroseal Corporation
  • AFC has manufactured freezing bags since 1982.
  • AFC bags are marketed by CellGenix GmbH.
  • CellGenix GmbH is exhibiting Afc bags at this
    conference in the exhibit area.
  • This notice is required by ISCT and the Medical
    Education Collaborative.

SOP for validation of freezing bags
  • The SOP presented here is on the ISCT website.
    It may be downloaded and used as a template for
    your validation work.
  • This SOP is also on my website WWW.TOAFC.COM
    under FAQ.
  • The download is in WORD, it does not have these
    powerpoint notes.

The SOP in ISO format
  • 1.0 Purpose
  • 2.0 Scope
  • 3.0 Responsibility
  • 4.0 References and Applicable Documents
  • 5.0 Materials and Equipment
  • 6.0 Health and Safety Considerations
  • 7.0 Documentation Requirements
  • 8.0 Definitions
  • 9.0 ProcessGeneral Description
  • 10.0 ProcessDetails
  • 11.0 Other Pertinent Information
  • 12.0 Training Requirements
  • 13.0 Revision History
  • 14.0 Attachments

Title block of SOP
  • Director of Quality Assurance ______________
    Date ______
  • Director of Cryopreservation ______________
    Date ______
  • Effective date _________________
  • Next Review date _________________

Where to go for guidance
Where do you get a validation protocol?
  • The test conditions for (validation) runs should
    encompass upper and lower processing limits and
    circumstances, including those within standard
    operating procedures, which pose the greatest
    chance of process or product failure compared to
    ideal conditions.
  • From Part IV, General Concepts, Guidelines on
    General Principles of Process Validation.

FDA definition of Validation Protocol
  • A written plan stating how validation will be
    conducted, including test parameters, product
    characteristics, production equipment, and
    decision points on what constitutes acceptable
    test results.
  • From Section III, Guideline on General Principles
    of Process Validation.

FDA Elements of Process Validation
  • Tests and challenges should be repeated a
    sufficient number of times to assure reliable and
    meaningful results.
  • From Section VII, Elements of Process Validation,
    paragraph 1. http//

1. Purpose
  • This is a Standard Operating Procedure for
    validating cryopreservation bags and the process
    used for freezing, while frozen and during
  • (Note In order to validate for cryopreservation,
    you must cryopreserve. This requires a
    Cryopreservation process.)

2. Scope.
  • SCOPE This Procedure includes selecting bags for
    validation, the validation process and the
    validation report. This procedure is used to
    determine whether the bag and can survive extreme
    conditions of processing. This will validate the
    bag, ports, overwrap, label, and all in process
    devices, procedures and components. Validation
    must challenge the limits of the procedure while
    using the same equipment, processes, and
    personnel that are used for routine processes.
    .. More.

2. .Scope
  • This procedure challenges the container by
    subjecting it to the lowest temperature that
    might be achieved in a laboratory setting. And
    the procedure challenges the integrity of the bag
    by both over filling and under filling. This
    procedure challenges the durability of the bag by
    extreme temperature shock. This procedure
    challenges the entire process by subjecting the
    product to ten sequential freeze thaw cycles.
    This procedure includes a form to report

Drop Test
  • Cant I do a simple test?

Use your own SOPs
  • Validation must use the processes that your
    institution employs, and the processes must test
    the bags at the worst case conditions of your
  • Note not the worst case you can think of.

Durability test (worst case)
How to validate
  • My bags sometimes fail when thawing because they
    inflate and crack.
  • Can I just use an inflation test?

Pressure Testing
  • Can I validate the bags by inflating them until
    they break?
  • Is inflation a part of your standard procedure?

Scope There are testing labs
  • Can they do the validation for me?

  • Can I send it out for validation?
  • Only if you send all your products to the same
    place for cryopreservation.

No, you cant send it out
  • Validation must be done by the same people who do
    the cryopreservation for the institution. They
    must use the same equipment and the same protocol
    that they use when freezing actual products.

So, what is validation?
  • Process validation is establishing documented
    evidence which provides a high degree of
    assurance that a specific process will
    consistently produce a product meeting its
    pre-determined specifications and quality
  • From Section IV. General Concepts, Guideline on
    General Principles of Process Validation.

3. Who is Responsible
  • RESPONSIBILITY Validation of cryopreservation
    bags is the responsibility of the Quality
    Assurance director who reports to the CEO of this
    institution. The Quality Assurance director must
    approve and sign this procedure. In the event
    that there is no Quality Assurance Director, a
    representative of management who reports to the
    chief executive officer or the chief executive
    officer may sign. The Cryopreservation
    Department Director also must approve and sign
    this procedure. It is the responsibility of the
    cryopreservation technical staff of this
    institution to carry out this procedure.

4. References
  • 21CFR820, sections A-H.
  • 21CFR820.75 Process Validation
  • 21CFR10.90 Guideline on General Principles of
    Process Validation.
  • 21CFR1271.230 Process Validation (a) General.
    Where the results of processing described in para
    1271.220 cannot be fully verified by subsequent
    inspection and tests, you must validate and
    approve the process according to established
  • FDAs Guide to Inspections of Quality Systems
    Manual QSIT, August 1999. Particularly Section
    Production and Process Controls, inspection
    point number 4 If the results of the process
    reviewed cannot be fully verified, confirm that
    the process was validated by reviewing the
    validation study. (Search the Web FDA QSIT).
  • This institutions SOPs relating to freezing of
  • NOTE ISO any standards?

ISO Standards?
  • I have not found ISO standards for
    cryopreservation or freezing.

What about this ISO standard?
  • (ISO 116072003(E) Packaging for terminally
    sterilized medical devices Is not applicable.)
  • (Search the web ISO CRYOPRESERVATION, or ISO

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5. Materials and Equipment
  • 5.1. Bags
  • 5.2 Overwraps
  • 5.3. Cryopreservation media
  • 5.4. Red food dye (unless the freeze media is
    deep red).
  • 5.5. Sealer
  • 5.6. Controlled Rate Freezer
  • 5.7. Freezer
  • 5.8. Thawing Water Bath, 40 degrees Celsius.

6. Health and Safety Considerations
  • 6.1. Handling frozen materials can cause frost
  • 6.2. Persons following this procedure must be
    trained in the safe use of cryopreservation
    equipment, freezers, controlled rate freezers,
    and water baths.

7. Documentation Requirements
  • 7.1. Validation requires a clear data set that
    documents the work done, the results of the work,
    the name of the person who did the work, and the
  • 7.2. Section 14 contains a worksheet for
    recording the results of this procedure. The
    worksheet must be signed and dated at the
    appropriate places.
  • 7.3. This SOP is maintained in the Quality
    Policy Manual and is reviewed at least annually,
    or whenever the materials or procedures change.
  • 7.4. The validation report is maintained in the
    Quality Manual under Quality Systems Records
    and is available for audit.

8. Definitions
  • 8.0. SOP shall mean Standard Operating
  • 8.1. Procedure and Process mean the same
    thing and are interchangeable.
  • 8.2. Critical equipment shall mean any
    equipment or process that meets the following
    criteria (a). Equipment that would make a
    defective product if it were not use properly.
    (b). Equipment that has adjustments or settings
    that control its operation. (c). Equipment that
    would create a defect that would not be detected
    by visual observation or subsequent procedures.
  • 8.3. Cryopreservation shall mean freezing to
    some temperature below the solidification of the
    product. ...

8. .Definitions
  • 8.4. Filling shall mean adding fluid to the bag
    to a volume indicated by this institutions SOPs.
    If not otherwise specified filling will be to a
    thickness of 1.0 centimeters.
  • 8.5. Temperature and volume measurements shall be
    made in the International Metric units.
  • 8.6. Water Bath shall mean a container of water
    in which the water is maintained at 40 degrees
  • 8.7. Product shall mean the bag, its contents,
    and any necessary accessories.
  • 8.8. Overwrap shall mean a bag or other
    covering that is placed over the product to wrap
    the product bag, contain any leaks, and protect
    the product from contamination.
  • 8.9. this institution shall mean the
    organization manufacturing the product and doing
    the validation.
  • 8.10. Cryopreservation media shall mean the
    fluids that normally would be used to fill the
    bag, not necessarily including any biologics.

9. Process General Description
  • Records for Critical Equipment are examined to
    ensure that the equipment is installed in a
    quality manner (IQ). Records for critical
    equipment are examined to ensure that it operates
    in a quality manner (OQ). Records for critical
    equipment are examined to see that proper
    maintenance has been done. Each of these
    records IQ, OQ, and maintenance are recorded.
    Obtain and reference any instructions from the
    bag manufacturer regarding use of the bag.

9. Process General Description
  • Ten bags are filled using this institutions
    cryopreservation media following the SOPs of this
    institution. The fill solution is supplemented
    with red food dye to permit easy visualization of
    any leak. The bags are frozen in a controlled
    rate freezer following this institutions SOPs.
    The bags are transferred into liquid nitrogen
    overnight or longer as noted in the validation
    report. The bags are removed from liquid
    nitrogen and placed directly into a 40 degree
    water bath. The bags are observed for leaks into
    the water bath as evidenced by escape of red dye,
    readability of labels, and condition of ports and
    overwrap. Then the freeze thaw cycle is repeated
    nine more times with the same bags. The
    observations are recorded on the attached form.

10. Process Details
  • Obtain and review the IQ, OQ, and maintenance
    records for the controlled rate freezer and
    record the file numbers on the Validation Record
  • Obtain and review the IQ, OQ, and maintenance
    records for the freezer and record the file
    numbers on the Validation Record form.
  • Obtain and review the IQ, OQ, and maintenance
    records for the water bath and record the file
    numbers on the Validation Record form.
  • Select nine bags to be tested These are taken
    from the top, middle, and bottom of the next 3
    cases of product bags that would normally be
  • Record the manufacturer and lot numbers of the
  • Mark the bags with sequential numbers using this
    institutions method.

10. Process - Details
  • Add red food dye to the cryopreservation media to
    develop a deep red color (if the media is not
    already deep red).
  • Fill bags 1, 4, and 7 with the volume of fluid
    that this institution recommends.
  • Fill bags 2, 5, and 8 with twice the volume of
    fluid that this institution recommends.
  • Fill bags 3, 6, and 9 with half the volume of
    fluid that this institution recommends.
  • Freeze the bags in the controlled rate freezer to
    -80 degrees Celsius according to this
    institutions procedure.
  • Transfer the bags to the liquid phase of a liquid
    nitrogen freezer.
  • Store overnight or over weekend.
  • Remove bags from the freezer and place directly
    into 40 degree Celsius water bath.
  • Observe for leakage of fluid. Report any leakage
    as leak.
  • Observe for loss of protective cover over port.
    Report cover loss as cover lost
  • Observe for ability to read bag identity
    information. Report inability to read as loss
    of identity.
  • Observe for breaks in the overwrap. Report as
    overwrap failure.
  • Sign and date each entry.
  • Remove any bags that leak or otherwise fail from
    the validation cycle.
  • Repeat the freeze and thaw process for 10 cycles.
  • Submit to the Quality Assurance Director for
  • Any failure invalidates the process.

11. Other Pertinent Information
  • In this section, include any information that
    relates to the validation process such as
    references that are not directly used. For
    example literature on freezing.
  • In this section, include previous or other
    pertinent validation studies or notes.

12. Training Requirements
  • Technicians responsible for validation work shall
    read and be familiar with 21CFR820.
  • Technicians responsible for validation shall
    complete Cryopreservation training as required by
    this institution.
  • Quality Assurance shall train technicians in this
    procedure and shall properly note training in the
    employees training record.

13. Revision History
  • This revision (A) is given as an example of how
    to make and record a revision to the procedure.
  • Food Color dye was added to the procedure. The
    reason for the change is that it was noted that
    after several freezes, the dye in the
    cryopreservation media becomes pale and harder to
    see. Changes were made to section 5, 9, and 10.
    No other changes were made. The revision level
    was changed from 0 to A. Changes made by John
    Jones, April 4, 2006.

  • Freezer IQ record number _______
  • Freezer OQ record number _______
  • Freezer Maintenance record number _______
  • Controlled rate freezer IQ _______
  • Controlled rate freezer OQ _______
  • Controlled rate freezer Maintenance record
    number _________
  • Water Bath IQ record number _______
  • Water Bath OQ record number _______
  • Water Bath Maintenance record number ___________
  • Bag Manufacturer _______________________________
  • Bag Lot Numbers ________________________________
  • Training record of person doing Validation
    procedure __________
  • This is the data taken of 10 freeze/thaw cycles
  • Cycle results
    Date Signed
  • 1. _______________ _____
  • 2. _______________ _____
  • 3. _______________ _____
  • 4. _______________ _____

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