Background and Aim: This study sought to explore International Islamic University Malaysia (IIUM) third-year pharmacy students’ knowledge and attitudes regarding immunization using a questionnaire. Method: A crosssectional prospective survey was carried out among 89 students to obtain demographic data. Other data were collected using knowledge and attitude immunization questionnaires consisting of twenty questions related to knowledge and five questions related to attitude and practice. The final questionnaire was subsequently discussed among specialist pharmacists who are experts in this field and who judged the face and content validity of the questionnaire. Descriptive statistics were used and internal consistency was tested using Cronbach’s alpha coefficient. Results: Employing the recommended scoring method, the mean+standard deviation of the knowledge and attitude scores was 11.7614 ± 3.30445. Cronbach’s alpha test for internal consistency was 0.582 for the 25 items in the knowledge and attitude questionnaire. Conclusion: The study concluded that the knowledge and attitude immunization questionnaire appears to be reliable and valid for measuring knowledge, attitude, and practices among third-year pharmacy students at IIUM. It can be used in future research as it was suitable to determine the knowledge and attitudes regarding immunization among pharmacy students.