人血清蛋白-葡聚糖偶合抗原的制备与表征

Preparation and Characterization of Conjugated Antigen between Human Serum Albumin and Dextrans

将葡聚糖用NaIO4氧化成带醛基的产物,再与人血清蛋白(HSA)中的氨基反应形成不稳定的席夫碱,在还原剂硼氰氢化钠(NaCNBH3)的作用下,成为稳定的拟糖蛋白抗原,并对影响偶合反应的几个方面进行讨论,包括:不同分子质量的葡聚糖、葡聚糖氧化所用NaIO4的量、氧化葡聚糖与HSA的比例和偶合时间。结果表明:偶合反应的最佳条件为:葡聚糖T40与氧化剂NaIO4物质的量比为1:187,偶合溶液pH9.0,氧化葡聚糖T40与HSA物质的量比为4:1,偶合时间12h。质量相等的不同类型葡聚糖,葡聚糖分子质量越大,氧化时需要的氧化剂越多,氧化度越高接枝度也越高。

Dextrans were oxidized using sodium periodate（NaIO4） to obtain products with aldehyde groups,which further reacted with amino（-NH2） groups of human serum albumin（HSA） to form unstable Schiff＇s base,and stable neoglycoprotein was then obtained in the presence of NaCNBH3 as reductant.In this study,the conjugation between human serum albumin and dextrans was investigated with respect to the effects of dextran type,amount of NaIO4 used for dextran oxidization,ratio between oxidized selected dextran and HSA,and reaction time.The optimum reaction conditions for the conjugation between human serum albumin and dextrans were found to be：dextran T40/NaIO4 molar ratio 1：187,environmental pH 9.0,and oxidized dextran T40/HSA molar ratio 4：1 for a reaction duration of 12 h.In addition,larger amounts of NaIO4 were required for the oxidation of dextrans with larger molecular weight;the higher degree of oxidation（DO）,the higher degree of grafting（DG）.