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抗HSV_1单克隆抗体阿糖胞苷偶联物制备条件的分步优化

Fractional optimization experiment of production condition for anti-HSV_1 monoclonal antibody conjugate and its determination
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摘要 目的 :建立高效液相凝胶过滤色谱法测定偶联物的偶联率、中间产物的生成量 ;通过制备条件分步优化实验来提高偶联率。方法 :制备条件从投料比、pH、温度、反应时间等方面来逐项优化反应条件以期能提高偶联率 ;检测方法采用高效液相凝胶过滤色谱法 ,以 0 .0 2mol·L-1 pH6 .8磷酸盐缓冲溶液为流动相 ,流速为 0 .5mL·min-1 ,检测波长为 2 80nm。结果 :通过建立高效液相凝胶过滤色谱法来指导优化实验 ,摸索得到新的投料比和 pH ,使偶联率从1 0 .1 9%提高到 1 3.4 8%。结论 :本实验建立的高效液相凝胶过滤色谱法简便、快速、灵敏、准确 ,对优化实验有指导意义 ,使偶联率得到了提高。 OBJECTIVE To establish a gel filtration chromatography (GFC) for determination of conjugation rate and yield of intermediate; conjugation rate was increased through optimation experiment. METHODS In order to increase output,the material ratio for the reaction, pH, temperature and reaction time were adjusted, GFC was applied to quantitative analysis, mobile phase was 0.02 mol·L -1 pH 6.8 dihydogenphosphate solution with a flow rate of 0.5 mL·min -1, the UV detection wavelength was 280 nm. RESULTS The conjugation rate increased from 10.19% to 13.48%. CONCLUSION The established method is simple,rapid, sensitive and accurate for optimizing experimental condition and increasing the conjugation rate.
出处 《中国医院药学杂志》 CAS CSCD 北大核心 2004年第5期266-268,共3页 Chinese Journal of Hospital Pharmacy
关键词 抗HSV1单克隆抗体 阿糖胞苷 优化实验 凝胶过滤色谱 偶联率 anti-HSV_1 monoclonal antibody cytarabine optimization experiment gel filtration chromatography conjugation rate
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