PREPARATION AND PROPERTIES OF FLUORESCENT POLYSACCHARIDES

PREPARATION AND PROPERTIES OF FLUORESCENT POLYSACCHARIDES
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DOI:
10.1016/0003-2697(83)90590-0
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发表时间:
1983-01-01
影响因子:
2.9
通讯作者:
ROSEN, SD
ROSEN, SD
中科院分区:
生物学4区
文献类型:
--
作者:
GLABE, CG;HARTY, PK;ROSEN, SD

文献摘要

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报道了一种制备多糖荧光素衍生物的新方法。这些衍生物通过用溴化氰活化多糖并随后与荧光素胺反应来制备。确定了偶联反应的最佳条件。使用该程序,制备了多种多糖的荧光素衍生物。取代度在3.0 × 10 - 6范围内。10-3到2.4倍。10-2得到荧光素摩尔数/单糖当量摩尔数。荧光衍生物是稳定的:在22 ℃温育后没有检测到游离荧光素。48小时或在-10 ℃下进行。C为4个月。荧光素衍生的多糖在抑制凝集素介导的血凝中具有与未衍生的多糖相同的效力。此外,由于125 I掺入荧光素中,这些荧光多糖可以被放射性碘标记至超过106 dpm/μ g的比活度。125 I-岩藻糖苷和125 I-肝素的细胞结合特性与相应的未衍生化多糖没有区别。这种制备荧光多糖的一般方法应该产生用于定位和定量细胞表面碳水化合物结合蛋白(凝集素)的有用试剂。
A new method for preparing fluorescein derivatives of polysaccharides is described. These derivatives are prepared by activation of the polysaccharide with cyanogen bromide and subsequent reaction with fluoresceinamine. The optimum conditions for coupling were established. Using this procedure, fluorescein derivatives of a wide variety of polysaccharides were prepared. Degrees of substitution in the range of 3.0 .times. 10-3 to 2.4 .times. 10-2 mol of fluorescein/mol of monosaccharide equivalent were obtained. The fluorescent derivatives are stable: no free fluorescein was detected after incubation at 22.degree. C for 48 h or at -10.degree. C for 4 mo. The fluorescein-derivatized polysaccharides had the same potency in inhibiting lectin-mediated hemagglutination as the underivatized polysaccharide. In addition, these fluorescent polysaccharides can be radioiodinated to specific activities exceeding of 106 dpm/.mu.g due to incorporation of 125I into fluorescein. The cell binding properties of 125I-fucoidin and 125I-heparin are indistinguishable from the corresponding underivatized polysaccharides. This general approach for preparing fluorescent polysaccharides should produce useful reagents for localizing and quantifying cell surface carbohydrate-binding proteins (lectins).