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应用 GP5 蛋白研制抗猪繁殖与呼吸综合征病毒的单克隆抗体。

Application of GP5 protein to develop monoclonal antibody against porcine reproductive and respiratory syndrome virus.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.

出版信息

Virol Sin. 2011 Aug;26(4):267-72. doi: 10.1007/s12250-011-3198-5. Epub 2011 Aug 17.

Abstract

In this study, a panel of monoclonal antibodies (mAbs) against Porcine reproductive and respiratory syndrome virus(PRRSV), named as 8C9 and4B4, were produced by fusing SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with the PRRSV (TCID(50)=5.5), screened by the indirect ELISA and subjected to several limiting dilutions. mAbs were then identified by biological characterization. Among the two fusion cell strains, 8C9 belonged to the IgG1 subclass and 4B4 belonged to the IgG2a subclass. The titers in cell culture supernatant and abdomen liquor reached to 1:10(4)and 1:10(5), respectively. The specificity test indicated that the two cells had specific reactions for the PRRSV and GP5 protein respectively, and no reaction with Classical swine fever virus (CSFV) or Swine vesicular disease virus (SVDV). The molecular weights of the heavy chain and light chain were about 45.0 kDa and 25.0 kDa, respectively. In neutralization activity tests, the results showed that the prepared mAb 4B4 can protect 50% of cells with no CPE in dilution up to 1:512, but mAB 8C9 has no neutralization activities to PRRSV.

摘要

在这项研究中,通过融合 SP2/0 骨髓瘤细胞和用 PRRSV(TCID(50)= 5.5)免疫的 BALB/c 小鼠的脾细胞,产生了针对猪繁殖与呼吸综合征病毒(PRRSV)的单克隆抗体(mAbs),命名为 8C9 和 4B4,通过间接 ELISA 筛选,并进行了几次有限稀释。mAbs 然后通过生物学特性进行鉴定。在两种融合细胞株中,8C9 属于 IgG1 亚类,4B4 属于 IgG2a 亚类。细胞培养上清液和腹水中的滴度分别达到 1:10(4)和 1:10(5)。特异性试验表明,两种细胞分别对 PRRSV 和 GP5 蛋白具有特异性反应,与猪瘟病毒(CSFV)或猪水疱病病毒(SVDV)无反应。重链和轻链的分子量分别约为 45.0 kDa 和 25.0 kDa。在中和活性试验中,结果表明,制备的 mAb 4B4 可在不产生 CPE 的情况下保护 50%的细胞,稀释度高达 1:512,但 mAb 8C9 对 PRRSV 没有中和活性。

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