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猪链球菌2型人源分离株MRP单克隆
抗体的制备与鉴定
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王海丽1，徐公义1，王长军2，唐家琪2*
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(1.聊城职业技术学院，山东 聊城252000；2.南京军区军事医学研究所，江苏 南京210002)
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摘要：
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为建立一种针对猪链球菌2型（SS2）毒力因子的特异性检测方法，研制了抗SS2溶菌酶释放蛋白（MRP）单克隆抗体（McAb），并对其生物学活性进行了分析。将表达、纯化的SS2菌株MRP免疫BALB/c小鼠，采用淋巴细胞杂交瘤技术将免疫鼠脾细胞与SP2/0骨髓瘤细胞融合，以纯化的MRP蛋白作为包被抗原，通过间接ELISA方法筛选获得了6株能稳定分泌抗MRP McAb的杂交瘤细胞系2B8、3G5、1G1、3G9、1B6和2C3。经用间接ELISA方法测定，单抗腹水的抗体效价均超过1∶105，杂交瘤细胞培养上清液的效价为1∶128～1∶1024。Western-blotting鉴定结果表明，6株单抗均具有特异的生物学活性；以2B8单抗腹水和SS2多克隆抗血清建立的夹心ELISA方法可特异地检测MRP。 |
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关键词：
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猪链球菌2型；溶菌酶释放蛋白；单克隆抗体；鉴定 |
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中图分类号：S 852.611：Q 813.2 文献标识码：
A 文章编号：1673-4696(2008)05-0422-04 |
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Preparation and identification of monoclonal antibodies against
muramidase released protein(MRP) of Streptococcus suis
serotype 2 isolated from human |
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WANG Hai-li1,XU Gong-yi1,WANG
Chang-jun2,TANG Jia-qi2 |
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(1.Liaocheng Vocational and Technical College，Liaocheng 252000,China;
2.Research Institute for Military Medicine of Nanjing Military Command,Nanjing 210002,China） |
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Abstract:To establish a specific method for detection of toxicity factors of Streptococcus suis serotype 2(SS2),anti-MRP monoclonal antibodies (McAbs) were developed and their biological activities analyzed.The screening result of indirect ELISA with the purified MRP as coating antigen showed that six hybridoma cell lines 2B8,3G5,1G1,3G9,1B6 and 2C3 secreting McAb against MRP were obtained by fusion of mouse myeloma cells SP2/0 with splenic cells from BALB/c mice immunized with the purified expression product of MRP.The titers of cultivation supernatants of the 6 hybridoma cell lines ranged from 1∶128 to 1∶1024 and the titers of ascites from the mice were over 1∶105.The six McAbs were recognized specifi-cally by MRP in Western-blotting test.A sandwich ELISA for specific detection of MRP was developed using ascites from hybridoma cell line 2B8 and polyclonal anti-serum against SS2. |
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Key words: Streptococcus suis serotype 2;muramidase released protein(MRP);McAb;identification |
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