High-Throughput Carbohydrate Microarray Analysis of 24 Lectins

摘要

Lectins, non-immunoglobulin proteins that bind carbohydrates, play a central role in a wide range of biological processes such as cell-cell recognition, viral and bacterial pathogenesis, and inflammation.[1], [2] Moreover, they are used extensively as research tools, diagnostics, and therapeutics. For example, mistletoe lectin is in clinical trials as an anticancer agent.[3] Therefore, a fundamental understanding of carbohydrate-protein interactions and comprehensive information on lectin specificity is critical. Unfortunately, evaluation of lectin specificity is not trivial. One common method involves measuring the binding of lectins to cells, tissues, and glycoproteins. This approach frequently uncovers interesting and useful binding properties, however, cells, tissues, and glycoproteins display complex mixtures of carbohydrate epitopes. Therefore, it is exceedingly difficult to determine the specific carbohydrate structures being recognized by the lectin. An alternative approach involves measuring binding to structurally defined carbohydrate epitopes through techniques such as isothermal calorimetry (ITC), mono- and oligosaccharide inhibition studies, enzyme-linked lectin assays (ELLA), and surface plasmon resonance assays (SPR). Unfortunately, these methods can be labor intensive, require large amounts of carbohydrates, and/or be difficult to perform in a high-throughput fashion. Moreover, these studies have typically been limited to the small number of carbohydrate epitopes that were readily accessible. Although lectin specificity has been studied often, much more comprehensive information is still needed.