液相色谱-ELISA测定结合疫苗中游离蛋白含量可行性的初步探讨  

The primary study on the feasibility of determining the free carrier protein concentration of conjugate vaccines with LC-ELISA method

作　　者：谭小梅[1] 杨朝晖[1] 周园[1] 赵志强[1] 张萍[1] 刘月萍[1] 计永莉[1] 杜琳[1] 谢贵林[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2008年第1期43-47,共5页Progress In Microbiology and Immunology

摘　　要：用适宜分离范围的色谱柱,预先确定细菌多糖蛋白结合疫苗载体蛋白的洗脱峰出峰时间;然后将细菌多糖-蛋白结合疫苗用相同条件进行分离,分部收集各洗脱峰并直接包被酶标板,与适宜浓度的载体蛋白特异抗体反应后,加入酶标二抗测定各洗脱峰样品中载体蛋白的吸光值;把载体蛋白出峰后的吸光值总和计为游离载体蛋白质吸光值;以游离载体蛋白吸光值占所有洗脱峰吸光值的比例计算游离载体蛋白含量。以伤寒Vi结合疫苗(载体蛋白rEPA)和甲型副伤寒结合疫苗(载体蛋白为TT)作为实验材料探讨方法的可行性。结果说明,该方法具有较好的特异性和敏感度。To develop a new method for determining the free carrier protein concentration of conjugate vaccines, the free carrier protein was separated from the protein-polysaccharlde conjugate through liquid chromatography. Each fraction of elution peak was collected as the sample, which coated on ELISA plates directly, then the A450 of each well was determined, and the curve was protracted compared to the sample serial number. The free protein concentration was calculated according to the corresponding peak proportion. Typhi Vi-rEPA conjugate vaccine and Paratyphi A O-SP-Tr conjugate vaccine were analyzed, the results showed that this method has feasibility to determine the free carrier protein concentration of conjugate vaccines.

关 键 词：结合疫苗 游离蛋白质 ELISA 液相色谱 

分 类 号：R392[医药卫生—免疫学]