Phytopathology. 2000 Feb;90(2):183-90. doi: 10.1094/PHYTO.2000.90.2.183.
ABSTRACT A panel of recombinant single-chain antibodies (scFvs) against structural proteins of Tomato spotted wilt virus (TSWV) was retrieved from a human combinatorial scFv antibody library using the novel phage display technique. After subcloning the encoding DNA sequences in the expression vector pSKAP/S, which allowed the scFvs to be expressed as alkaline phosphatase fusion proteins, 17 different scFv antibodies were obtained. Of these, 12 scFvs were directed against the nucleoprotein (N) and 5, putatively, against the glycoproteins (G1 and G2). Five of the N-specific antibodies cross-reacted with two other tospoviruses (Tomato chlorotic spot virus and Groundnut ringspot virus), but none recognized the more distantly related tospoviruses Impatiens necrotic spot virus, Watermelon silverleaf mottle virus, Iris yellow spot virus, or Physalis severe mottle virus. The successful use of one of the antibodies as coating and detection reagent in a double-antibody sandwich enzyme-linked immunosorbent assay showed the potential of the phage display system in obtaining antibodies for routine TSWV diagnosis.
摘要 利用噬菌体展示技术从人源组合 scFv 抗体文库中筛选出了一组针对番茄斑萎病毒（TSWV）结构蛋白的重组单链抗体（scFv）。将编码 DNA 序列亚克隆到表达载体 pSKAP/S 中后，scFv 可以表达为碱性磷酸酶融合蛋白，共获得了 17 种不同的 scFv 抗体。其中 12 种 scFv 针对核蛋白（N），推测 5 种针对糖蛋白（G1 和 G2）。5 种 N 特异性抗体与另外两种 TSWV（番茄斑萎病毒和花生环斑病毒）发生交叉反应，但没有一种抗体识别亲缘关系较远的 TSWV，如矮牵牛坏死斑点病毒、西瓜银叶斑点病毒、鸢尾黄斑点病毒或辣椒严重斑驳病毒。其中一种抗体成功地用作双抗体夹心酶联免疫吸附试验中的包被和检测试剂，这表明噬菌体展示系统在获得常规 TSWV 诊断抗体方面具有潜力。
