Carbohydrates are often found linked to lipids or proteins, to form glycoconjugates such as glycolipids, glycoproteins and proteoglycans. These glycoconjugates play important biological roles, involving cell-cell recognition, inflammation, immune response, tumor metastasis and in viral/bacterial/parasitic infections. However, it is difficult to obtain these naturally occurring glycoconjugates as they are often heterogeneous compositions, which causes batch-to-batch variability of structure and activity often leading an incomplete understanding of their mechanism of action. Thus, the efficient preparation of synthetic glycoconjugates, possessing well-defined compositions and reproducible biological properties is highly desirable. In the present review, we summarize the advances in site-selective approaches for glycoconjugate synthesis and, in particular, the preparation of polysaccharide vaccines.