Significant strides have been achieved in unraveling the 3D structures of proteins; however, the structural investigation of glycoproteins presents distinctive challenges due to the unpredictable and heterogeneous nature of protein glycosylation, coupled with the intricate and flexible structures of glycoprotein glycans. Conventional bacterial expression systems are inadequate for producing glycoproteins, posing a hurdle for structural biology endeavors. Confronting these obstacles, nuclear magnetic resonance (NMR) spectroscopy, in synergy with various analytical and preparative techniques, stands out as a potent resolution. Recombinant glycoproteins can now be expressed using stable isotope labeling through various eukaryotic production platforms. High-performance liquid chromatography mapping and mass spectrometry enable the analysis of glycoforms, while genetic engineering of production vehicles and in vitro enzymatic reactions facilitate glycoprotein glycan remodeling. Stable-isotope-assisted NMR techniques offer a robust approach, furnishing detailed insights into the conformational dynamics and interactions of carbohydrate chains in solution. This information provides a valuable understanding of the functional mechanisms of glycoprotein glycans. Moreover, the synergy of NMR spectroscopy and computational approaches empowers us to enhance the functionality of carbohydrate chains by designing and optimizing their conformational spaces. This strategic approach lays the structural foundation for crafting glycoproteins with tailored, and even artificial, glycoforms.