火箭免疫电泳在W135群脑膜炎球菌发酵工艺优化中的应用  

Application of rocket immunoelectrophoresis in the optimization of group W135 meningococcus fermentation process

作　　者：许博 杨千苇 侯亚莉 于飞飞 刘月萍 应连芳 Xu Bo;Yang Qianwei;Hou Yali;Yu Feifei;Liu Yueping;Ying Lianfang(Quality Control Department,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 201403,China;Pilot Plant Test Lab,Lanzhou Institute of Biological Products Co.,Ltd.,Lanzhou 730046,China;First Department of Research,Lanzhou Institute ofBiological Products Co.,Ltd.,Lanzhou 730046,China)

机构地区：[1]上海生物制品研究所有限责任公司质量检定室,201403 [2]兰州生物制品研究所有限责任公司中试研究室,730046 [3]兰州生物制品研究所有限责任公司第一研究室,730046

出　　处：《国际生物制品学杂志》2019年第5期227-230,共4页International Journal of Biologicals

摘　　要：目的应用火箭免疫电泳(rocket immunoelectrophoresis,RIEP)检测W135群脑膜炎球菌(group W135 meningococcus,MenW135)发酵液的多糖含量,来指导MenW135发酵工艺的改进。方法制备家兔MenW135抗血清,并确定RIEP琼脂糖凝胶中的抗血清浓度。分别以不同发酵条件培养MenW135,并分别在培养2、3、4、5、6 h时取样,用分光光度法检测菌液的吸光度(A),用RIEP检测菌液的多糖含量。结果检测MenW135多糖的RIEP琼脂糖凝胶中的抗血清浓度确定为0.1%。在MenW135发酵2~5 h期间,MenW135多糖含量与其A值呈正相关性。然而,在MenW135发酵5~6 h期间,菌液的A值不增加,但菌液的多糖含量仍在升高。结论相对于通过菌液的A值来观察菌液的多糖含量,RIEP检测能直接获得菌液的多糖含量,从而更好地指导MenW135多糖疫苗发酵工艺的优化。Objective To apply rocket immunoelectrophoresis(RIEP)to detecting the polysaccharide content of W135 group meningococcus(MenW135)fermentation broth,in order to guide the process optimization of MenW135 fermentation.Methods The rabbits anti-MenW135 antiserum was prepared,and the concentration of the antiserum in RIEP agarose gel was determined.MenW135 was cultured under different fermentation conditions,and samples were taken at 2,3,4,5,and 6 h,respectively.The absorbance(A)of the bacteria broth was measured by spectrophotometry,and the polysaccharide content of the bacteria broth was detected by RIEP.Results The antiserum concentration in the RIEP agarose gel for detecting the MenW135 polysaccharide was determined to be 0.1%.During the first 2-5 h of bacteria fermentation,the polysaccharide content of MenW135 was positively correlated with its A value.However,during 5-6 h of bacteria fermentation,the A value of bacteria broth did not increase,while polysaccharide content of bacteria broth still increased.Conclusion Compared to using the A value of bacteria broth to observe polysaccharide content of bacteria broth,RIEP detection can directly obtain the polysaccharide content,which can guide the fermentation process optimization of MenW135 polysaccharide vaccine better.

关 键 词：免疫电泳 奈瑟球菌脑膜炎 多糖类 分光光度法 

分 类 号：R28[医药卫生—中药学]