Generation and characterisation of recombinant FMDV antibodies: Applications for advancing diagnostic and laboratory assays.
Generation and characterisation of recombinant FMDV antibodies: Applications for advancing diagnostic and laboratory assays.
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DOI：
10.1371/journal.pone.0201853
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Jackson T
中科院分区：
文献类型：
--
作者：
Shimmon G;Kotecha A;Ren J;Asfor AS;Newman J;Berryman S;Cottam EM;Gold S;Tuthill TJ;King DP;Brocchi E;King AMQ;Owens R;Fry EE;Stuart DI;Burman A;Jackson T
口蹄疫（FMD）影响经济上重要的牲畜，是最具传染性的病毒性疾病之一。最常用的FMD诊断测定是夹心ELISA。然而，这种ELISA的主要缺点是它需要在小动物中产生的抗FMD病毒（FMDV）亚型特异性抗体。使用抗FMDV单克隆抗体（MAbs）作为检测试剂可以部分地克服这个问题。然而，单克隆抗体的长期使用可能存在问题，可能需要更换。在此，我们使用识别O型FMDV的小鼠MAb D9（MAb D9）的Fv（片段可变）区构建了嵌合抗体（小鼠/兔D9）和Fab（片段抗原结合）（小鼠/牛D9）。小鼠/兔D9嵌合抗体保留了MAb D9的FMDV亚型特异性，并且在FMDV检测ELISA以及常规实验室测定中表现良好。冷冻电子显微镜分析证实了与抗原位点1的接合，肽竞争研究将残基VP 1 147处的天冬氨酸鉴定为D9表位的新组分。这种嵌合表达方法是保存有价值的FMDV抗体的简单但有效的方法，并且具有在重组系统中无限产生抗体和抗体片段的潜力，同时对3R（替换、减少和细化）原则产生积极影响。
Foot-and-mouth disease (FMD) affects economically important livestock and is one of the most contagious viral diseases. The most commonly used FMD diagnostic assay is a sandwich ELISA. However, the main disadvantage of this ELISA is that it requires anti-FMD virus (FMDV) serotype-specific antibodies raised in small animals. This problem can be, in part, overcome by using anti-FMDV monoclonal antibodies (MAbs) as detecting reagents. However, the long-term use of MAbs may be problematic and they may need to be replaced. Here we have constructed chimeric antibodies (mouse/rabbit D9) and Fabs (fragment antigen-binding) (mouse/cattle D9) using the Fv (fragment variable) regions of a mouse MAb, D9 (MAb D9), which recognises type O FMDV. The mouse/rabbit D9 chimeric antibody retained the FMDV serotype-specificity of MAb D9 and performed well in a FMDV detection ELISA as well as in routine laboratory assays. Cryo-electron microscopy analysis confirmed engagement with antigenic site 1 and peptide competition studies identified the aspartic acid at residue VP1 147 as a novel component of the D9 epitope. This chimeric expression approach is a simple but effective way to preserve valuable FMDV antibodies, and has the potential for unlimited generation of antibodies and antibody fragments in recombinant systems with the concomitant positive impacts on the 3Rs (Replacement, Reduction and Refinement) principles.
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影响因子：
14.9
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