14型肺炎球菌荚膜多糖-破伤风类毒素结合物中游离多糖的测定方法  

Determination of free polysaccharides in pneumococcal polysaccharide serotype 14-tetanus toxoid conjugates

作　　者：姚静[1] 廖磊[1] 黄林[1] 周觉非 廖红梅[1] 刘威[1] 

机构地区：[1]成都生物制品研究所有限责任公司,成都610023

出　　处：《中国新药杂志》2012年第10期1127-1130,共4页Chinese Journal of New Drugs

摘　　要：目的:建立一种简单有效的测定肺炎球菌荚膜多糖-破伤风类毒素结合物中游离多糖的方法。方法:根据脱氧胆酸(DOC)在酸性条件下可快速将蛋白质沉淀的原理,采用不同pH值的1%DOC溶液分别对14型肺炎球菌荚膜多糖(Ps14)、破伤风类毒素(TT)及Ps14-TT结合物进行处理,观察不同pH值1%DOC溶液对它们的影响,来确定最佳反应pH值。随后采用精密度试验和准确度试验对该方法进行验证。结果:Ps14-TT结合物经1%DOC处理后,在酸性条件下(1 mol.L-1HCl),结合物及蛋白质被完全沉淀,而未结合的Ps14存在于上清液中,采用蒽酮法测定结合物当中的游离多糖含量。结论:该方法重复性,稳定性,精密度,准确度均达到检测要求,适用于检测Ps14-TT结合物中游离多糖的测定。Objective： To establish a simple and effective assay for determining the concentrations of free ysa OC charides in pneumococcal polysaccharide-tetanus toxoid conjugates. Methods： Since deoxycholic acid can precipitate protein in acidic conditions, pneumococcal polysaccharide Psl4, tetanus toxoid and Psl4- conjugates were treated with deferent pH values 1% DOC solution. The impact of different pH values of 1% DOC solution on them was observed, and the optimal reaction pH was determined. Then this method with precision test and accuracy test were verified. Results： After Psl4-TT conjugates were treated with 1% DOC solution, the conjugates and free protein were completely precipitated in acidic conditions （1 mol· L-1 HC1）, with free polysac- charides in the supernatant. The concentration of free polysaccharides then was determined using anthrone assay Conclusion： The method shows good reproducibility, stability, precision and accuracy, and meets the detection requirements. It is applicable to detect the concentration of free polysaccharides in Psl4-TT

关 键 词：多糖蛋白结合物 破伤风类毒素 游离多糖 脱氧胆酸 

分 类 号：R186[医药卫生—流行病学]