This study investigated the effects of an online interprofessional education (IPE) program on nursing students at a university in Japan. It conducted a comparative analysis between traditional face-to-face and online interventions. All students who enrolled in the “Team Medical Practice” course in both 2019 and 2020 were invited to participate. After team-based learning with different healthcare professions, we quantitatively analyzed pre- and post-intervention outcomes using two measures: the Readiness for Interprofessional Learning Scale (RIPLS) and the IPE Questionnaire TSUKUBA model (IPET). We compared the results of 153 nursing students. The RIPLS results demonstrate significant pre- and post-intervention score differences in the teamwork and collaboration subscale for the online group only. The post-IPE score analysis revealed that scores were significantly higher in the online group in all subscales: Teamwork and collaboration, Opportunities for IPE, and Uniqueness of profession. Based on the IPET results, there were no significant differences in pre- and post-intervention scores. However, the online group showed a significant increase in post-intervention scores in participation in group work, thoughts about the team in health and welfare, and thoughts about interprofessional collaboration. These findings indicate that the online IPE program improved nursing students’ readiness for interprofessional learning, demonstrating practical efficacy.