GTP Scenario # 2 September 17,2005

Scenario # 2 Dr. Good received IND approval for CD8 depletion of allogeneic PBMC for a Phase I/II clinical trial. Dr. Good received IND approval for CD8 depletion of allogeneic PBMC for a Phase I/II clinical trial.

Laboratory Information Cell Therapy Laboratory: Cell Therapy Laboratory: Unclassified facility. Unclassified facility. BSC cleaned with 10% bleach before use (1 product per BSC). BSC cleaned with 10% bleach before use (1 product per BSC). BSC is certified annually. BSC is certified annually. Depletion is performed in the general laboratory area (not within BSC). Depletion is performed in the general laboratory area (not within BSC). Product sampling is performed in the BSC. Product sampling is performed in the BSC. What level of EM and Cleaning is required? What level of EM and Cleaning is required?

What should be considered? Evaluate the conditions. Evaluate the conditions. The process is ‘functionally closed’. The process is ‘functionally closed’. The product is a 351 IND product. The product is a 351 IND product. The laboratory in which the depletion process occurs is an unclassified room. The laboratory in which the depletion process occurs is an unclassified room. Other processing (sampling, antibody addition) occurs in the BSC. Other processing (sampling, antibody addition) occurs in the BSC.

Product Monitoring Sterility: Pre & Post processing? Sterility: Pre & Post processing? Recommended Recommended Periodic audit review of sterility results? Periodic audit review of sterility results? Recommended Recommended

BSC Environmental Monitoring Settle plates during processing? Settle plates during processing? Recommend as part of process validation. Determine need based on data. Recommend as part of process validation. Determine need based on data. Surface sampling after cleaning? Surface sampling after cleaning? Perform as part of validation and as periodic check of cleaning procedure. Perform as part of validation and as periodic check of cleaning procedure. Airborne sampling during processing? Airborne sampling during processing? Not recommended. Not recommended.

Facility Monitoring Perform air particle counts during processing? Perform air particle counts during processing? Recommend monthly checks during laboratory ‘working hours’ (dynamic monitoring). Recommend monthly checks during laboratory ‘working hours’ (dynamic monitoring).

Personnel Monitoring Perform on day of processing after the staff completes the process? Perform on day of processing after the staff completes the process? Recommend performing during processing only if products begin to have positive sterility results. Recommend performing during processing only if products begin to have positive sterility results.

EM & PM Result Review How often? How often? Recommend monthly review. Recommend monthly review. Does this need to be done before product release? Does this need to be done before product release? Review cannot be part of product release; the product is infused immediately after processing (fresh). Review cannot be part of product release; the product is infused immediately after processing (fresh).

Cleaning BSC? BSC? Depletion Device? Depletion Device? Lab benchtops? Lab benchtops? Lab floors? Lab floors? Document Review? Document Review?

Cleaning Recommendations BSC BSC Cleaning with 10% bleach before & after use. Cleaning with 10% bleach before & after use. Unless post-processing cultures become positive (investigation outcome). Unless post-processing cultures become positive (investigation outcome).

Cleaning Recommendations Depletion Device & Benchtops: Depletion Device & Benchtops: Clean before & after use; Device using manufacturer’s instructions, Benchtop using 70% IPA. Clean before & after use; Device using manufacturer’s instructions, Benchtop using 70% IPA. Floors: Floors: Clean on a periodic basis. Clean on a periodic basis. Unless post-processing cultures become positive (investigation outcome). Unless post-processing cultures become positive (investigation outcome). Unless there is a spill. Unless there is a spill.

Cleaning Document Review How often? How often? Recommend periodic document review. Recommend periodic document review. Does this need to be done before product release? Does this need to be done before product release? Review should not be part of product release. Review should not be part of product release.

Variation # 1 Would you do anything differently if the blood had to be ficolled in conical tubes before depletion? Would you do anything differently if the blood had to be ficolled in conical tubes before depletion? Yes, refer to ‘open’ process recommendations. Yes, refer to ‘open’ process recommendations.

Variation # 2 Would you change anything if this were a Phase III study? Would you change anything if this were a Phase III study? No, unless required by the FDA. No, unless required by the FDA.