Abstract: Glycoconjugate vaccines are essential tools for preventing bacterial infectious diseases. Currently licensed glycoconjugate vaccines are produced through a stepwise process that involves the extraction of native polysaccharide antigens, purification of carrier proteins, and subsequent in vitro chemical conjugation. In recent years, the rapid advancement of emerging technologies such as synthetic biology has driven the development of innovative strategies for designing and manufacturing glycoconjugate vaccines. This review highlights progress in two representative technologies: protein glycan coupling technology (PGCT) and cell-free glycoprotein synthesis (CFGpS). It also briefly explores additional emerging approaches in glycoconjugate vaccine development, including generalized modules for membrane antigens (GMMA), semi-synthetic glycoconjugate vaccines, non-covalent coupling strategies, and click chemistry-based conjugation using non-natural amino acids. Looking ahead, advancements in artificial intelligence (AI) are expected to accelerate the computational design and engineering of oligosac-charyltransferases and carrier proteins, ultimately improving the immunogenicity and manufacturing efficiency of next-generation glycoconjugate vaccines.