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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。
Predetermined peak elution time of the carrier protein of the bacterial polysaccharide protein conjugate vaccine with a column of a suitable separation range; and then separating the bacterial polysaccharide-protein conjugate vaccine under the same conditions, fractionally collecting the elution peaks and directly coating ELISA plate, and the appropriate concentration of the carrier protein-specific antibody reaction, the enzyme was added to determine the secondary antibody peak elution peak carrier protein absorbance values; the carrier protein peak after the sum of absorbance as the free carrier protein absorbance value The free carrier protein content was calculated as the ratio of the absorbance of free carrier protein to the absorbance of all eluting peaks. The feasibility of the method was discussed with the combination of typhoid Vi combined vaccine (carrier protein rEPA) and paratyphoid A (TT as carrier protein) as experimental materials. The results show that the method has good specificity and sensitivity.