Yamaguchi T, Iwata K, Kobayashi M, Ogawa M, Fukushi H, Hirai K
Department of Veterinary Microbiology, Faculty of Agriculture, Gifu University, Japan.
Arch Virol. 1996;141(8):1493-507. doi: 10.1007/BF01718250.
Twenty hybridoma cell lines producing monoclonal antibodies (MAbs) against serotype 1 infectious bursal disease virus (IBDV) of GBF-1 and the attenuated GBF-1E strains were produced. The MAbs recognized major structural proteins VP2 and VP3. MAb recognition sites were mapped using recombinant Escherichia coli clones which expressed N-terminal and (or) C-terminal truncated virus antigens, and competitive-binding assays. At least 3 conformation-dependent serotype 1 specific virus neutralizing antigenic sites and a linear antigenic site were defined on VP2 and VP3, respectively. Two of the conformational virus neutralizing antigenic sites were localized in the central area of VP2 consisting of 156 amino acid residues, and the linear epitope was localized in C-terminal 105 amino acid residues of VP3. Another conformational virus neutralizing antigenic site recognized with the virus neutralizing MAb GK-5 was not defined because GK-5 did not react with virus antigen expressed in recombinant E. coli. The conformational antigenic site was supposed to be composed of tertiary or quaternary protein structure, which may not be constructed in recombinant E. coli.
制备了20株杂交瘤细胞系,它们产生针对GBF - 1和减毒GBF - 1E株血清型1传染性法氏囊病病毒(IBDV)的单克隆抗体(MAb)。这些单克隆抗体识别主要结构蛋白VP2和VP3。使用表达N端和(或)C端截短病毒抗原的重组大肠杆菌克隆以及竞争结合试验来定位单克隆抗体识别位点。在VP2和VP3上分别定义了至少3个构象依赖性血清型1特异性病毒中和抗原位点和1个线性抗原位点。其中2个构象性病毒中和抗原位点位于由156个氨基酸残基组成的VP2中央区域,线性表位位于VP3的C端105个氨基酸残基处。另一个用病毒中和单克隆抗体GK - 5识别的构象性病毒中和抗原位点未明确,因为GK - 5不与重组大肠杆菌中表达的病毒抗原发生反应。该构象性抗原位点推测由蛋白质的三级或四级结构组成,而在重组大肠杆菌中可能无法构建。