Mislovicová D, Masárová J, Svitel J, Mendichi R, Soltés L, Gemeiner P, Danielsson B
Institute of Chemistry, Slovak Academy of Sciences, SK-842 38 Bratislava, Slovakia.
Bioconjug Chem. 2002 Jan-Feb;13(1):136-42. doi: 10.1021/bc015517u.
Neoglycoconjugates were prepared from mannan isolated from yeast Saccharomyces cerevisiae and activated by periodate oxidation to create aldehyde groups. Various degrees of oxidation introduced 11-28 aldehyde groups per mannan molecule and simultaneously resulted in a molar mass decrease from 46 to 44.5-31 kDa. The activated mannans were subsequently conjugated with bovine serum albumin forming neoglycoconjugates. Some parameters of these mannan-bovine serum albumin conjugates were characterized: saccharide content 25-30% w/w, molar mass within the range 169-246 kDa, and polydispersion (M(w)/M(n)) from 2.8 to 3.6. The interaction of these conjugates with lectin concanavalin A was studied using three different methods: (i) quantitative precipitation in solution; (ii) sorption to concanavalin A immobilized on bead cellulose; and (iii) kinetic measurement of the interaction by surface plasmon resonance. Quantitative precipitation assay showed only negligible differences in the precipitation course of original mannan and the corresponding mannan-bovine serum albumin conjugates. Both the sorption method (equilibrium method) and the surface plasmon resonance measurement (kinetic method) demonstrates that the values of dissociation constant K(D) of all synthetic neoglycoconjugates were within the range 10(-7) - 10(-8) mol x L(-1) (close to K(D) = 10(-8) mol x L(-1) determined by the sorption method for the original mannan). In conclusion, characterization of synthetic neoglycoconjugates confirmed that the method used for their preparation retained the ability of mannan moiety to interact with concanavalin A.
新糖缀合物是由从酿酒酵母中分离得到的甘露聚糖制备而成，通过高碘酸盐氧化将其活化以产生醛基。不同程度的氧化作用使得每个甘露聚糖分子引入了11 - 28个醛基，同时导致摩尔质量从46 kDa降至44.5 - 31 kDa。随后，活化的甘露聚糖与牛血清白蛋白结合形成新糖缀合物。对这些甘露聚糖 - 牛血清白蛋白缀合物的一些参数进行了表征：糖类含量为25 - 30% w/w，摩尔质量在169 - 246 kDa范围内，多分散性（M(w)/M(n)）为2.8至3.6。使用三种不同方法研究了这些缀合物与凝集素伴刀豆球蛋白A的相互作用：（i）溶液中的定量沉淀；（ii）吸附到固定在珠状纤维素上的伴刀豆球蛋白A；（iii）通过表面等离子体共振对相互作用进行动力学测量。定量沉淀分析表明，原始甘露聚糖和相应的甘露聚糖 - 牛血清白蛋白缀合物的沉淀过程仅存在可忽略不计的差异。吸附法（平衡法）和表面等离子体共振测量（动力学法）均表明，所有合成新糖缀合物的解离常数K(D)值均在10(-7) - 10(-8) mol·L(-1)范围内（接近通过吸附法测定的原始甘露聚糖的K(D) = 10(-8) mol·L(-1)）。总之，合成新糖缀合物的表征证实了用于其制备的方法保留了甘露聚糖部分与伴刀豆球蛋白A相互作用的能力。
