The introduction of vaccines for the treatment and prevention of bacterial or viral diseases in the early 19th century marked a crucial turning point in medical history. Since then, extensive immunization campaigns have eradicated smallpox and drastically reduced the number of diphtheria, tetanus, pertussis and measles cases worldwide. Although a broad selection of vaccines is available, there remains a need to develop additional vaccine candidates against a range of dangerous infectious diseases, preferably based on precise syntheses that lead to homogenous formulations. Different strategies for the construction of this type of vaccine candidates are being pursued. Glycoconjugate vaccines are successful in the fight against bacterial and viral infectious diseases. However, their exact mechanism of action remains largely unknown and the large-scale production of chemically defined constructs is challenging. In particular, the conjugation of the carbohydrate antigen to the protein carrier has proved to be crucial for the properties of these vaccines. This review highlights some of the latest findings and developments in the construction of glycoconjugate vaccines by means of site-specific chemical reactions.