Development of sugar chain-binding single-chain variable fragment antibody to adult T-cell leukemia cells using glyco-nanotechnology and phage display method
Development of sugar chain-binding single-chain variable fragment antibody to adult T-cell leukemia cells using glyco-nanotechnology and phage display method
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DOI：
10.1093/jb/mvy005
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发表时间：
2018-04-01
影响因子：
2.7
通讯作者：
Suda, Yasuo
中科院分区：
文献类型：
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作者：
Muchima, Kaname;Todaka, Taro;Suda, Yasuo
成人t细胞白血病（Adult T-cell leukemia， ATL）是由人t细胞白血病病毒1型感染引起的一种难治性血癌，需要有效的药物治疗。众所周知，细胞表面糖链的结构和表达水平取决于细胞的状态，如炎症和癌症。因此，ATL细胞表面糖链特异性抗体有望成为一种有效的诊断工具和抗ATL药物开发的有力候选物。在此，我们开发了一种稳定的糖链结合单链可变片段抗体（scFv），它可以通过纤维型糖芯片和噬菌体展示方法与ATL细胞结合。利用ATL细胞株释放的o聚糖制备纤维型糖片。然后利用纤维型Sugar Chips筛选来自人B细胞的显示scFv的噬菌体（多样性：1.04 × 10(8)），获得o -聚糖结合的scFv。流式细胞术分析显示，scFv主要与ATL细胞株结合。利用阵列式糖芯片固定化合成的糖胺聚糖双糖结构文库，评价了scFv的糖链结合特性。高度硫酸化的双糖结构对scFv有很高的亲和力。
Adult T-cell leukemia (ATL) is an intractable blood cancer caused by the infection of human T-cell leukemia virus type-1, and effective medical treatment is required. It is known that the structure and expression levels of cell surface sugar chains vary depending on cell states such as inflammation and cancer. Thus, it is expected that the antibody specific for ATL cell surface sugar chain would be an effective diagnostic tool and a strong candidate for the development of an anti-ATL drug. Here, we developed a stable sugar chainbinding single-chain variable fragment antibody (scFv) that can bind to ATL cells using a fibre-type Sugar Chip and phage display method. The fiber-type Sugar Chips were prepared using O-glycans released from ATL cell lines. The scFv-displaying phages derived from human B cells (diversity: 1.04 x 10(8)) were then screened using the fiber-type Sugar Chips, and an O-glycan-binding scFv was obtained. The flow cytometry analysis revealed that the scFv predominantly bound to ATL cell lines. The sugar chain-binding properties of the scFv was evaluated by array-type Sugar Chip immobilized with a library of synthetic glycosaminoglycan disaccharide structures. Highly sulphated disaccharide structures were found to have high affinity to scFv.