Objective: The objective of this study was to analyze the 100 most cited studies on vaccine. Methods: A comprehensive search of studies on vaccine was performed in the Web of Science Core Collection without year or language restrictions. The 100 top-cited studies were retrieved after screening abstracts or full-texts. The outcomes of bibliometric analysis included citation time, citation density, journal name, impact factor, publication year, article type, category, open access, and country of origin. Results: The citation times for the 100 top-cited studies ranged from 593 to 2406, with a median citation times of 834. The 100 top-cited studies were published in 32 journals, and the journal with the most studies was New England Journal of Medicine (n = 20). They were published between 1969 and 2012, and 4 authors published at least 2 studies as the first author. The USA contributed the most studies (n = 70), followed by Switzerland (n = 4), England (n = 4) and Finland (n = 4). Eighty-one studies were published as Article, while 19 were Review. Eleven studies were about vaccine for therapeutic and 68 studies were about vaccine for prophylactic. Conclusions: This is the first bibliometric analysis to provide a detailed list of the 100 most-cited studies on vaccine and helps to recognize the quality of the works, discoveries, and trends in the field.