Abstract Body: Background: Our system, comprised of 21 Northeast Ohio hospitals includes 3 Stroke Ready, 10 Primary and 1 Comprehensive Stroke Center (CSC). Alteplase was the standardized thrombolytic until 2022. Clinical trials support the efficacy and safety of Tenecteplase for the treatment of acute ischemic stroke (AIS). Tenecteplase administration is efficient with less room for error and is cost-effective. The transition from Alteplase to Tenecteplase for AIS patients eligible for thrombolysis occurred on June 6, 2022. Implementation transpired over 9 months. Purpose: To evaluate a health system’s experience transitioning to a novel thrombolytic. Methods: Stroke program and pharmacy stakeholders reviewed literature and agreed upon Tenecteplase 0.25mg/kg. The legal department approved as our new standard of care for AIS. Alteplase remained on formulary for pediatric stroke and other diagnoses. Guidelines and references were updated. A dosing table was created for rounding accuracy. Nurses were provided with dosing table and care instruction badge backers. Due to Tenecteplase package labeling for MI, supplies were repackaged into “TNK Kits”. Kits include a dosing table, visual step-by-step reconstitution instructions, 10mL syringe, 5mL syringe, 2 flush syringes and medication vials. EHR Alteplase references were changed to “thrombolysis”. Tenecteplase Stroke and MI order sets with auto-calculations were developed for safety. All impacted disciplines received education, including nursing skills demonstration. A retrospective review of system Alteplase administrations during 6/2020- 5/2022 was compared to Tenecteplase administrations throughout 6/2022- 5/2024. Results: Thrombolytic utilization increased from 464 patients (10.3%) treated with Alteplase versus 521 patients (12.3%) with Tenecteplase. The percentage of door to thrombolysis <30 minutes improved from 23.3% to 34.8% and <45 minutes from 60.4% to 73.2%. Door to puncture times for CSC direct arrivals that received Alteplase had a 93 minute median and CSC direct arrivals given Tenecteplase had a median of 82 minutes. Door to reperfusion median decreased by 23 minutes. The system Alteplase hemorrhagic complication rate was 2.8% compared to 2.5% for Tenecteplase. Current Alteplase wholesale cost is $10,560 and Tenecteplase is $8,854. Hence, a cost-savings of >$500,000 annually. Conclusion: Our system’s experience was successful in our goal to achieve high quality patient outcomes and improved care efficiency.