Miltenyi Biotec CryMACS Freezing Bag Validation

Transcription

1 VALIDATION PLAN Miltenyi Biotec CryMACS Freezing Bag Validation Model #: N/A Serial #: N/A (if not applicable, N/A) Added to Validation Plan Binder Table of Contents as In Progress by: : Validation Type Equipment Process Supplies and Reagents Prospective Concurrent Re-validation* Retrospective *If re-validation, describe reason for re-validation and proposed steps that will be completed: Purpose: Baxter Healthcare will no longer be supplying Cryocyte Freeze bags. Miltenyi Biotec has provided us CryoMACS Freeze bags which are very similar in look and size to the Cyrocyte Freeze bags and have recently been approved by the FDA. Miltenyi did an extensive validation on these bags using the CD34-depleted negative fraction from a CliniMACS CD34+ purified product. They did a comparison study against three other approved cyro-freeze bags using this CD34-depleted product. They looked at the following, post thaw: viable WBC by 7AAD, CD34% as a % of prefreeze content, CD34/L, CFU-GM and BFU-E. They also tested the bags durability by putting them through multiple integrity tests. Since this product has been validated by the company so extensively, a small in-house validation will be performed to ensure that it functions as well as the Cyrocyte bag using our protocols. Phase I of the validation is to ensure that the freeze bag chains can be created in a way that the tubing seals do not become compromised and that product, when distributed into multiple bags, is equal. Fresh product recovery will also be looked at during this phase. Phase II will examine at least three HPC, Cord Blood products both pre and post thaw. Each product will be cryopreserved in a Cryocyte bag and a CryoMACS bag. These two bags will be compared and the following data will be obtained: viable TNC recovery, viability, Total CD34 recovery, and CFC functional assay results. The bags will also be examined post LN2 storage to ensure that the bag did not become compromised during the controlledrate freeze process. Completion of this validation will show that the CryoMACS bag is equal to or better than the Cryocyte bag maintaining product sterility, viability and recovery. Equipment Installation, Installation and/or Operational Qualification Performance Qualification Study plan author Study plan approval Data review Final approval and implementation RESPONSIBILITIES N/A ACCEPTABLE RESULTS For this study the following minimal measurement parameters are required: Precision: Phase 1: Product should drain equally between the CryoMACS bags attached to the freeze chain. This should be reproducible a minimum of three times. Percent recovery of product should not be statistically different between the products examined. Cryobag Validation Template 2 Page 1 of 5

2 Phase 2: All results, post thaw comparison of the Cryocyte bag and the CryoMACS bag, should not be statistically different. This should be true for a minimum of three HPC, Cord Blood products. Accuracy: Post Thaw TNC recovery, viability, and Total CD34 recovery should not be more than ± 10% of those results found using the Cryocyte bag. Functional data from the CFC assay should also be statistically comparable between the two bag types Specificity: N/A Sensitivity: N/A Linearity and Range: Values may vary (Phase II) due to how the product is mixed and dispersed into the cryobags. More cells may be found in one bag compared to the other however viability should not vary drastically between the bags. Correlation should be acceptable (greater than R 2 ) for % viable cell recovery and functional parameters when comparing data sets from both products. PROCEDURE SOPs Needed- CP:11- Peripheral Blood HPC Cryopreservation CP:12 Cord Blood Cryopreservation CP:34 - Flow Cytometry - Acquisition and Analysis of HPC Products CP:18 - Cell Viability-Manual Cell Counts CP:62 - Sysmex XE 5000 Automated Hematology Analyzer Operation CP:21 - Colony-Forming Cell Assay CP:33- Cord Blood Thaw and Dilution Name and location of needed manuals AXP AutoXpress(TM) Platform Operator and Maintenance Manual Study forms needed- (indicate location or refer to relevant SOP) CryoMACS Phase 1 Worksheet Cord Blood Thaw and Dextran Dilution Worksheet HPC(C) Processing Worksheet Colony Forming Plating Assay Worksheet Other Equipment or Items Needed: See all Reagents and Special Supplies Required in SOPs CP:11, CP:12, CP:34, CP:18, CP:62, CP:21, and CP:33 QNS Cord Blood Units 250mL CryoMACS Freezing bags from Milteny Biotec Procedure: Keep all instrument printouts and attach to this validation A. Phase 1 1. Create a four bag cryopreservation harness using the CryoMACS Freeze bags a. Ensure the seal is intact and will not break 2. Obtain a QNS cord blood unit and QS it to ~140 ml with Plasmalyte A 3. Mix the product well then remove a 0.5 ml sample and analyzed on the AcT Diff 2 4. Attach the cord blood unit to the four bag harness and drain product into the four bags a. Drain product according to CP:11 5. Heat seal each bag then weigh each bag to determine the approximate volume drained 6. Mix each bag well then remove a small sample from each bag and analyze on the AcT Diff 2 7. Pool each bag into a single bag then mix the product well 8. Remove a small sample from the pooled product and analyze on the AcT Diff 2 then reweigh the bag b. Record initial TNC per bag (post drain / distribution) 9. Rinse each CryoMACS bag with ~10 ml of Plasmalyte A, add each rinse to the pooled product 10. Mix the pooled product bag then remove a small sample and run it on the AcT Diff Reweigh pooled product b. Record TNC per bag (post recovery from bag) 12. Calculate the percent recovery for the following: Cryobag Validation Template 2 Page 2 of 5

3 a. Product drained into CryoMACS bags (step 6) b. Pooled product volume and total TNC (step 8) c. Pooled product and recovered TNC with rinse (step 10) 13. Repeat all steps a minimum of two additional times B. Phase 2 1. Throughout procedure label each bag and tube with the following identifiers: a. Unique Product Identifier (example Product 1) b. Cryopreserved HPC, Cord Blood c. CryoMACS Validation d. 2. Obtain a QNS HPC, Cord Blood unit 3. Obtain a buffy coat from the unit by processing it using the AXP a. Follow the protocol in the AXP AutoXpress(TM) Platform Operator and Maintenance Manual, Chapter 5 pages 5-1 through 5-49 b. Complete the HPC(C) Processing Worksheet while processing 1) Record the lot number and expiration date of the AXP processing bag as well as any key supplies or reagents 2) Record the AXP serial number as well as any critical equipment that could affect the testing results include the serial/ctf# (i.e. incubator, Partec, Sysmex, ect.) 4. Add 37 ml of expressed plasma to the product 5. Mix the final product well then remove the following samples in the BSC: a. 1 ml for counts, viability, flow and CFC Assay 1) While in the BSC remove 100 µl for CFC Assay and place in a red top tube(ensure this sample does not leave the BSC) 6. Perform counts and viability according to the applicable SOP 7. Mix the product well then remove 28 ml and place it into a Cryocyte bag, remove the remaining 28 ml place it into a CryoMACS bag a. This step should be performed by the same individual for each product validated 8. Prepare the product for freezing and freeze according to CP:12 a. Use 7 ml of Dextran /DMSO freeze media for each cryobag b. Ensure product bags are labeled as stated above c. Record the lot numbers and expiration dates of the following supplies on the HPC(C) Processing worksheet: 1) Cryocyte Bag 2) CyroMACS Bag 3) Freeze Media d. List all critical equipment used that could affect the testing results, include the serial/ctf# (i.e. Cryomed, syringe pump) 9. While the product is freezing run a CD34 auto panel on the flow sample according to applicable SOP 10. Prepare and plate the CFC assay a. Complete the Colony Forming Plating Assay Worksheet 11. Place all product bags in quarantine temporary storage for a minimum of 24 hours prior to next step 12. Thaw and dilute each product separately according to SOP CP:33 a. Complete the Cord Blood Thaw and Dextran Dilution Worksheet for each bag b. Complete all testing listed in SOP CP: Repeat steps 1-12 for a minimum of two more products SYSTEM DESCRIPTION Expected results or function- Phase 1 percent recovery should not be statistically different between the products tested. Phase 2 post thaw viability, viable TNC recovery, and Total CD34 between the two bags should not vary more than ± 10%. CFC Assay results both pre and post thaw should be comparable and the results between the two bags for each product should not be statistically different. Critical Control Points- If expected ranges for recovery are not met, all circumstances related to product and product processing will be evaluated including their potential effect on recovery and viability. If necessary, additional product will be processed. Cryobag Validation Template 2 Page 3 of 5

4 Key Elements- Each product must be mixed well prior to sampling and prior to splitting the product pre-freeze. If the product is not adequately mixed prior to splitting it into the cryo-bags, the results may differ by more than 10 %. To eliminate technologist variability, one technologist will perform this step for each product involved in this validation. TARGET REPLICATES Each phase will be completed with a minimum of three cord blood products. Based on the recovery data obtained, the Lab Director may request further product studies. Any changes to this procedure will be documented in the execution write up. PLAN AUTHORIZATION SIGNATURES Study plan author Quality Assurance Plan Approval Laboratory Director Plan Approval Cryobag Validation Template 2 Page 4 of 5

5 PLAN EXECUTION Model #: Serial #: (if not applicable, N/A) Inclusive s of Study - From: Raw Data: Forms/Tables: Statistical Analysis: Graphs: To: DATA Results and Interpretation Statement(s) of comparisons and outcome: Conclusion Overall Conclusion of study results: AUTHORIZATION SIGNATURES Plan Execution Performed By: Quality Assurance Execution Approval Laboratory Director Execution Approval Release for Implementation This item is considered Validated Not Validated If Not Validated give reason: Required additional studies or monitoring after implementation (N/A if none): Further instruction/information if Not Validated (N/A if Validated): SOP implementation or modification Work-form creation or modification Personnel Notification: SOP Name: Work-form Name: Venue: IMPLEMENTATION PLAN : : : Personnel Training: Method: : Effective s Implementation of New Method: Cessation of existing Method: Cryobag Validation Template 2 Page 5 of 5