Synthetic multivalent glycans are widely employed as research tools for probing MLGI mechanisms as well as potential therapeutic interventions against specific MLGIs. When a multivalent glycan's spatial and orientation arrangement match those of a lectin's multiple binding sites, they will engage in simultaneous multivalent binding and form small 1:1 lectin-glycan complexes. This provides a great affinity and specificity enhancement and, hence, results in effective intervention. Whereas, those that do not have perfect spatial matches may inter-cross-link with multimeric lectins and form large scale lectin-glycan complexes. This often gives rise to a relatively low affinity enhancement and less effective results. However, information on how MLGI binding modes affect the affinity and underlying binding thermodynamics and kinetics remain largely underexplored. Due to the complex, multimeric, and often flexible nature of MLGIs, it has been difficult to reveal the exact binding modes of MLGIs by conventional biophysical techniques. For example, isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR) are two of the most widely employed biophysical tools for studying MLGI thermodynamics and kinetics. However, cross-linking interactions (widely observed in MLGIs) can make it difficult to interpret the ITC data. Moreover, ITC has relatively low sensitivity, and hence requires a large amount of sample, limiting its use for precious, difficult to obtain glycans. It is also difficult to dissect how individual LGIs determine the overall MLGI affinity and specificity by SPR as the density and orientation of surface immobilized binding partners can strongly affect the binding properties. As a result, these conventional biophysical techniques can only provide some, but not the whole set, of key biophysical parameters (e.g. binding thermodynamics, kinetics, binding modes, and binding site orientation) that are useful for both fundamental studies and therapeutic developments against specific MLGIs.