Chicken recombinant antibodies specific for very virulent infectious bursal disease virus
Chicken recombinant antibodies specific for very virulent infectious bursal disease virus
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针对高毒力传染性法氏囊病病毒的鸡重组抗体
DOI：
10.1007/s00705-006-0729-8
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发表时间：
2006
影响因子：
2.7
通讯作者：
J. Ignjatovic
中科院分区：
文献类型：
--
作者：
S. Sapats;L. Trinidad;G. Gould;Hans G. Heine;T. P. V. D. Berg;Nicolas Eterradossi;D. Jackwood;L. Parede;D. Toquín;J. Ignjatovic
以传染性法氏囊病病毒(VvIBDV)超强毒株CS89免疫鸡的免疫脾细胞为材料，构建了噬菌体展示单链可变区(ScFv)抗体库。对捕获的CS89病毒进行了3轮淘洗，文库由约9.2×107个克隆组成。对单个克隆的核苷酸测序分析表明，在ELISA中至少有22个独有的单链抗体与vvIBDV结合。对来自英国、中国、法国、比利时、非洲、巴西、印度尼西亚和荷兰的经典毒株、变异毒株或疫苗株以及多种vvIBDV分离株的单链抗体进行检测，鉴定出一种针对vvIBDV的抗体，命名为鸡重组抗体88(CREB 88)。Crab 88能够识别所有测试的vvIBDV毒株，无论其来源如何，并且与经典毒株、变异毒株和疫苗毒株没有反应，支持将该单链抗体用作区分vvIBDV毒株的有力诊断工具。免疫沉淀研究表明，螃蟹88针对的是位于主要中和蛋白VP2内的一个高构象表位。对IBDV VP2高变区的序列分析表明，Ile(256)和Ile(294)可能在与螃蟹88的结合中发挥作用。这是首个能够肯定区分vvIBDV和其他IBDV毒株的同类试剂。
A phage-displayed single chain variable fragment (scFv) antibody library was constructed from the immune spleen cells of chickens immunized with very virulent infectious bursal disease virus (vvIBDV) strain CS89. A library consisting of around 9.2 × 107clones was subjected to 3 rounds of panning against captured CS89 virus. Analysis of individual clones by nucleotide sequencing revealed at least 22 unique scFv antibodies binding to vvIBDV in ELISA. Testing of the scFv antibody panel in ELISA against classical, variant or vaccine strains and a wide variety of vvIBDV isolates from the UK, China, France, Belgium, Africa, Brazil, Indonesia and the Netherlands identified one antibody, termed chicken recombinant antibody 88 (CRAb 88) that was specific for vvIBDV. CRAb 88 was capable of recognizing all vvIBDV strains tested regardless of their country of origin and showed no reactivity with classical, variant or vaccine strains, lending support to the use of this scFv as a powerful diagnostic tool for the differentiation of vvIBDV strains. Immunoprecipitation studies revealed that CRAb 88 was directed towards a highly conformational epitope located within the major neutralizing protein VP2. Sequence analysis of the hypervariable region of VP2 of the IBDV strains tested indicate that Ile(256) and Ile(294) may play roles in binding of CRAb 88. This is the first reagent of its type capable of positively distinguishing vvIBDV from other IBDV strains.