Carbohydrate Microarray Profiling of Glycan-Binding Antibodies and Lectins

癌细胞在发病期间经历碳水化合物表达的急剧变化， 疾病的进展。因此，人们对理解为什么 这些变化的发生，它们如何导致疾病，并利用这些变化， 临床应用。碳水化合物结合抗体和凝集素在其中起着关键作用。 研究有几个原因。首先，抗体和凝集素被广泛用于检测和 监测碳水化合物表达的变化。在这种情况下，它们用于基本的 研究，诊断和治疗药物。全面了解他们的 特异性对于正确使用和解释结果至关重要。直到最近， 没有现成的资料。随着碳水化合物微阵列技术的出现， 我们现在可以快速分析这些蛋白质与各种碳水化合物的结合 抗原在过去，我们已经筛选了大量市售凝集素， 研究界使用的抗体试剂。我们发现，许多这些试剂不 显示预期的特异性。我们提供的关于特异性的信息将 能够更好地使用这些试剂，并便于解释结果。最近我们 一直在分析选定的抗体和凝集素与研究人员在合作， 德国癌症研究所，Abeome公司，和华盛顿大学圣路易斯分校在 在每种情况下，抗体和凝集素选择性地结合肿瘤并显示抗癌作用， 活动它们似乎与聚糖结合，但碳水化合物配体的身份是 未知该阵列已被用于分析这些蛋白质的结合特性， 识别其配体。此外，该阵列可用于开发新的抗体， 用于监测新的碳水化合物抗原的凝集素。该阵列在 开发结合GalN β 1 -3Gal抗原的单克隆抗体。使用这些 抗体，我们发现这种碳水化合物抗原改变了鳞状上皮细胞的表达， 细胞癌的子宫颈和表达的变化与预后相关。

Cancer cells undergo dramatic changes in carbohydrate expression during the onset and progression of disease. As a result, there has been considerable interest in understanding why these changes occur, how they contribute to the disease, and exploiting these changes for clinical use. Carbohydrate binding antibodies and lectins play a critical role in this research for several reasons. First, antibodies and lectins are used extensively to detect and monitor changes in carbohydrate expression. In this capacity, they are used for basic research, as diagnostics, and as therapeutic agents. A comprehensive understanding of their specificity is critical for proper use and interpretation of results. Until recently, this information was not readily available. With the advent of carbohydrate microarray technology, we can now rapidly analyze binding of these proteins to a wide variety of carbohydrate antigens. In the past, we have screened a large set of commercially available lectin and antibody reagents used in the research community. We found that many of these reagents did not display the expected specificity. The information we provided regarding specificity will enable better use of these reagents and facilitate interpretation of results. Recently, we have been profiling selected antibodies and lectins in collaborations with researchers at the German Cancer Research Institute, Abeome Corp., and University of Washington-St. Louis. In each case, the antibodies and lectins selectively bind tumors and display anti-cancer activity. They appeared to bind glycans, but the identities of the carbohydrate ligand(s) were unknown. The array has been used to profile the binding properties of these proteins and identify their ligands. In addition, the array can be used to develop new antibodies and lectins for monitoring new carbohydrate antigens. The array has been instrumental in the development of monoclonal antibodies that bind the GalNAca1-3Gal antigen. Using these antibodies, we have found that this carbohydrate antigen has altered expression on squamous cell carcinomas of the cervix and that changes in expression correlate with prognosis.