9. Attach a 2 inch 21G needle to a 3.0 mL syringe and draw 2.0 mL of air into the syringe. Inject 2.0 mL of air into the upright Leukine/Test Drug vial. DO NOT INVERT THE VIAL. Insert the 2 inch needle down to the bottom of the vial and slowly withdraw the entire contents of the vial making sure all liquid is removed. 10. Using the needle and syringe containing the Leukine/Test Drug from step #9, top-fill four 1.0 mL TB syringes with 0.4 mL of the Leukine/Test Drug in each syringe. 11. Attach a new sterile 3/8 inch 26G needle to each syringe containing 0.4 mL of Leukine/Test Drug. Appropriately label each syringe and dispense syringes for intradermal administration of study drug. The syringes must be used within 6 hours of mixing. CONFIDENTIAL Page 88 of 102 Protocol: HERCEPTIN® + E75/GM-CSF Version 3.1 ML25749 05 March 2017 Guidance for DTH Preparation from E75 Acetate Solution 1.5mg/1mL Vial To Prepare the DTH skin test, withdraw 0.07mL of the 1.5mg/1.0mL E75 acetate solution. The DTH skin test is administered intradermally using 0.07 mL (100 µg) of the E75 acetate solution. Supplies: All DTH skin test kits will contain an active vial, which contains 1.0 mL of E75 acetate in WFI at 1.5 mg/mL, Some DTH kits will contain a second vial which is a control vial containing 1.0 mL of Water For Injection (WFI). If the DTH kit contains the WFI, discard the WFI in the kit per your institution’s policy. E75 acetate solution preparation: 1. Remove cap from 1.5 mg/1mL E75 acetate solution and wipe with alcohol prep pad 2. Open 1 mL syringe with needle attached and uncap the needle 3. Draw > 0.07 mL of air into the syringe and inject into the E75 acetate solution vial 4. Withdraw 0.07 mL of E75 acetate solution into the syringe 5. Remove the syringe and the needle from the vial 6. Drawback syringe plunger to capture hub volume into syringe 7.