0 Dead. 110 Abbreviated Title: GBM Pembro HSPPC Version Date: 03/26/2024 16.2 TUMOR TISSUE SUBMISSION FOR V ACCINE MANUFACTURE See Appendix 16.3 for detailed instructions for handling, preparation, and submission of tumor tissue for HSPPC-96 vaccine manufacture. The Tumor Procurement Form (TPF) must be included with the tissue sent to Agenus Inc. Kits for shipping tissue will be supplied by Agenus and should be ordered at time of IRB approval because they will be needed at time of surgery: Supply and Logistics Agenus Inc. Primary Contact: Tel: +1 (781) 674-4400 x4486 (office) Email: supply&logistics@agenusbio.com The dimensions of the kits are 17" x 17" x 15". Please order one or two kits at a time, based on your anticipated use, and please allow 3-4 days for shipping. Upon receipt of a kit with patient tumor at Agenus, a new kit will automatically be replenished for the site. 111 Abbreviated Title: GBM Pembro HSPPC Version Date: 03/26/2024 16.3 VACCINE TISSUE P ROCUREMENT AND S HIPPING I NSTRUCTIONS Tissue procurement kits provided by Agenus Inc. include: o50-mL nonpyrogenic tubes and patient information labels oDry Ice Contents Sticker oAbsorbent paper oPlastic bags oOrange-capped pressure vessels (with foam insert) oCardboard pieces (to support base of pressure vessel and top of dry ice) oTumor Procurement Form (TPF) oShipping box and all shipping/address labels Sites must provide the dry ice (8kg). o The site should contact Agenus at supply&logistics@agenusbio.com to request courier documents (pre-paid shipping label for tumor shipment from site to Agenus) 1 day in advance of the desired shipment and the shipping label will be emailed to the site. The shipment of tissue to Agenus for vaccine creation should be delayed until confirmation of histologic diagnosis, MGMT status and IDH mutational status that would make the tumor eligible for vaccine production. Tumor specimens for vaccine production must be frozen and stored at -80°C ± 20°C. The tumor specimen must not arrive on the weekend or a holiday unless this has been agreed to in advance by Agenus. If the tumor specimen will not be shipped immediately, it should be placed into an -80° (+/- 20°) C freezer until ready to pack and ship. Do this within a maximum of 30 minutes of removal. For tumor specimen stored at sites prior to shipping to Agenus (i.e. overnight or for weekends/holidays), the freezer does not need to be sterile, but it is recommended that it be monitored for temperature (24/7) with alarms or other mechanism for identifying when temperature goes out of specification. The freezer or its location should have controlled access 112 Abbreviated Title: GBM Pembro HSPPC Version Date: 03/26/2024 (locked with specified group members having access). The freezer should also be on a preventative maintenance program and ideally be hooked up to a back-up power system. In the OR, wrap or place the tumor specimen in a sterile specimen cup, cloth or towel, and place in a basin, on ice (not dry ice). Transport the specimen immediately to the pathology/frozen section suite. Share the following steps with the surgical pathologist, along with the tissue procurement kit: a. Use basic sterile technique. Care should be taken to avoid any possible cross- contamination with tissues or fluids from other patients. b. As small a section of tumor as possible should be kept for routine pathologic studies and diagnosis. Tumor for routine pathologic diagnosis should be processed according to institutional standards. c. The pathologist should assess viability of the tumor. d. When there is necrosis and/or cystic degeneration in the tumor specimen, the necrotic tissue and/or cystic component should be removed and should not be sent to Agenus. e.All available tumor aside from what will be used for routine pathologic studies and diagnosis should be sent for HSPPC-96 vaccine preparation. A minimum of ≥ 7 grams of fresh, non-necrotic tumor should be sent (if <7 grams, may not produce sufficient vaccine). Tumor procurement and shipping procedure a. The pathologist should section the tissue into 1-2 cm2 sections. The sections should then be placed carefully, maintaining sterility, into the 50-mL nonpyrogenic tubes. Each tube should be filled no more than ½-⅔ full to allow for expansion of tumor during freezing. b. Each tube must be properly labeled (patient/subject identifiers are a match on all tubes). c. Each tube should be placed in a plastic pouch, and the pouch sealed closed. d. Wrap the tubes in bubble wrap or place in the foam insert in the upright position. e. Absorbent paper should be placed in the bottom of the orange capped pressure vessel. The sealed tubes in the foam insert are then placed into the pressure vessel 113 Abbreviated Title: GBM Pembro HSPPC Version Date: 03/26/2024 and the vessels lid is screwed shut. Dry ice should NOT be placed inside the pressure vessel. f. Place some dry ice in the bottom of the insulated shipper box. g. The pressure vessel should be placed above the dry ice, with the base seated in the cardboard support, and more dry ice packed surrounding the container. h. Dry ice should be added to about 10 cm from the top of the box, a second cardboard insert placed above the ice, and the foam plug placed into the box. If an insufficient amount of dry ice is placed in the box, and the tissue is not frozen on arrival, it cannot be processed and the patient must be withdrawn from the study. i.