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Background: Human Papillomavirus (HPV) vaccination rates remain low in the U.S., particularly among minorities and low-income patients. In 2014, a Mayo Clinic study of uninsured, low-income, working, Volunteer in Medicine (VIM) Clinic patients in Jacksonville, Florida found a statistically significant lower awareness of HPV (50.3% vs 63.6%) and the HPV vaccine (32.1% vs 62.7%) than the national population. As a result, our team collaborated with Merck to develop a free HPV vaccination program and video education campaign for VIM patients. Methods: We assessed VIM patients HPV vaccination knowledge, awareness, and prevalence of hesitancy towards receiving the vaccine. VIM patients (ages 18-45) with no previous history of HPV vaccination were invited to participate in this study. The Parents Attitudes about Childhood Vaccines (PACV) survey was used to evaluate vaccine hesitancy. Patients who declined the HPV vaccine were shown an educational video to assess effectiveness in persuading patients’ opinion. Each participant received a vaccine hesitancy PACV score, which was compared with their vaccine disposition. Mayo Clinic IRB approval of the study was obtained. Results: Our analysis included 43 patients (34 females, 9 males) with a median age of 40 (age range 27- 45). Patients had overall knowledge of HPV of 88.4%, with 69.8% having heard of the HPV vaccine. 85.7% PACV survey respondents were deemed non-hesitant (PACV scores 0-49), and 80.6% of the non-hesitant subset (29/36) agreed to undergo vaccination. The average PACV score of vaccine-interested participants was lower (25.2) than the average of participants not interested in the vaccine (36.7). Black participants had a statistically significant higher PACV score than White participants. The PACV scores of individuals who were already familiar with the HPV vaccine were significantly lower than those of participants who were unfamiliar with the HPV vaccine. Only 20% (2/10) of individuals, who had previously declined vaccination, agreed to receive the HPV vaccine after viewing the HPV educational video. Of the 33 participants who agreed to receive the free HPV vaccine, only 9 (27.3%) completed all three doses. Conclusions: HPV vaccine awareness and knowledge at VIM increased compared to the 2014 cohort. Vaccine hesitancy was not observed in most patients, yet video education was effective in only a small portion of patients with vaccine hesitancy. Additionally, a minority of patients who agreed to receive the free HPV vaccine series completed all three doses. Our results suggests that free HPV vaccination and video education is not sufficient to achieve adequate vaccination rates among low-income uninsured young adults, nor overcoming vaccine hesitancy. Innovative measures are warranted to improve vaccination rates in this population. Citation Format: Hiram Diaz, Gerardo Colon-Otero, Laura M. Pacheco-Spann, Christopher C. DeStephano, Sandra Casanova-Leyva. Assessing knowledge, vaccine hesitancy, and completion rates of free HPV vaccination in uninsured/underinsured low-income patients. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5539.