The problems of microbial infections and the emergence of drug-resistant microbes are increasingly serious, causing countless loss of lives and economic loss. The discovery and study of host defense peptides opened a new avenue in developing antimicrobial regents, and have attracted a lot of attention in recent years. Compared with natural host defense peptides, synthetic antimicrobial polypeptides can be conveniently synthesized in large scale and with low cost. Furthermore, saccharide-polypeptide conjugates have been valued for their optimal effect on antimicrobial properties and biocompatibility. In this review article, we provide an overview of the development and progress of antimicrobial polypeptides and saccharide-polypeptide conjugates regarding their structural design, biological functions and antimicrobial mechanism. By pointing out the challenges, we also provide future prospects of this research field from our perspectives.