Synthesis and biological properties of mannosylated starburst poly(amidoamine) dendrimers
Synthesis and biological properties of mannosylated starburst poly(amidoamine) dendrimers
复制标题
DOI：
10.1021/bc970126u
复制
发表时间：
1997-09-01
影响因子：
4.7
通讯作者：
Roy, R
中科院分区：
文献类型：
--
作者：
Page, D;Roy, R
以甘露吡喃糖苷残基为末端的星型PAMAM树枝状聚合物易于从其表面上具有高密度胺官能团的市售树枝状聚合物和对异硫氰酸基苯基2，3，4，6-四-O-乙酰基-α-D-甘露吡喃糖苷大规模且高产率地合成。使用酶联凝集素测定法（ELLA）和浊度分析，评价了具有多达32个甘露糖苷单元的这类新的单分散新糖缀合物的前四代作为来自伴刀豆球蛋白A（Con A）和豌豆（豌豆凝集素）的植物血凝素的配体。这些glycodendrimers，连同参考单糖的结合特性，确定使用酵母甘露聚糖作为包被抗原和过氧化物酶标记的凝集素。这些甘露糖基化树枝状聚合物被证明是有效的抑制剂，其IC 50值比作为标准的单体甲基α-D-吡喃甘露糖苷的IC 50值好400倍。它们的亲脂性足以在微量滴定板测定中直接用作包被抗原。此外，他们的能力，结合和形成不溶性碳水化合物凝集素复合物也证明了径向双免疫扩散和浊度分析。此外，使用聚丙烯酰胺凝胶电泳（SDS-PAGE）也证明了这些配体选择性地从粗凝集素混合物中沉淀甘露糖结合蛋白（Con A）的能力。这些多价的neoglycoconjugates被证明构成新的生物色谱材料的高亲和力，易于分离的碳水化合物结合蛋白。
Starburst PAMAM dendrimers ending with mannopyranoside residues were readily synthesized in large scale and good yields from commercially available dendrimers bearing high-density amine functionality on their surface and p-isothiocyanatophenyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranoside. The first four generations of this novel class of monodispersed neoglycoconjugates having up to 32 mannoside units were evaluated as ligands for the phytohemagglutinins from concanavalin A (Con A) and Pisum sativum (pea lectin) using enzyme-linked lectin assay (ELLA) and turbidimetric analyses. The binding properties of these glycodendrimers, together with reference monosaccharides, were determined using yeast mannan as a coating antigen and peroxidase-labeled lectins. These mannosylated dendrimers were demonstrated to be potent inhibitors with IC50 values 400 times better than those of monomeric methyl alpha-D-mannopyranoside taken as a standard. Their lipophilic character was shown to be sufficient for their direct use as coating antigens in microtiter plate assays. Moreover, their ability to bind and form insoluble carbohydrate-lectin complexes was also demonstrated by radial double immunodiffusion and turbidimetric analyses. Furthermore, the ability of these ligands to selectively precipitate a mannose-binding protein (Con A) from a crude lectin mixture was also demonstrated using polyacrylamide gel electrophoresis (SDS-PAGE). These multivalent neoglycoconjugates were shown to constitute novel biochromatography materials of high affinity for the easy isolation of carbohydrate-binding proteins.