摘要
硫酸多糖 (91 1 )还原性末端的半缩醛基 ,通过还原胺化反应与酪胺 (Tyr)的氨基共价偶联 ,91 1 -Tyr中酪胺引入的仲氨基通过与异硫氰酸酯荧光素 (FITC)进行亲核反应 ,实现对 91 1还原末端的选择性荧光标记 .用 UV-Vis吸收光谱、1 H NMR和 HPSEC对偶联与标记结果进行确证 ,从 1 H NMR谱推测 91 1与酪胺的偶联率及 FITC对 91 1 -Tyr标记率分别为 60 %和 80 % .由于采用的是 91 1末端选择性标记 ,对 91 1的抗凝活性无明显影响 ,也无明显的细胞毒性 .以荧光标记的 91 1作为探针 ,对淋巴细胞有较强的选择性标记染色 .
The reducing terminal of marine sulfated polysaccharide(911) was used to selectively insert primary and secondary amines by reductive amination. 911 was bounded to 4\|(2\|aminoethylphenol), followed by labeling the 911\|Tyr with fluorescein\|5\|isothiocyanate(FITC) at the secondary amino group. High\|performance size\|exclusion chromatography(HPSEC), ultraviolet/visible(UV\|Vis) spectroscopy and nuclear magnetic resonance spectroscopy(NMR) demonstrated the binding of tyramine to 911 and the labeling of FITC to 911\|Tyr respectively. Furthermore, \{\}\+1H NMR spectra and UV\|Vis spectra revealed about 60% binding of tyramine to 911 and about 80% labeling of FITC to 911\|Tyr. The anticoagulant activity of labeled fluorescent compound exhibited 13.40 U/mg equivalent to that of unlabled 911. In addition, the labeled compound was demonstrated to fluorescently stain lymphocytes without any cytotoxity. Taken together, the selectively 'end point attached' method might be widely used for fluorecent labeling to those polysaccharides with reducing terminal.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2002年第9期1704-1708,共5页
Chemical Journal of Chinese Universities
基金
国家自然科学基金 (批准号 :3 0 0 70 694)资助
