Chae J S, Choi J K, Lim H T, Cha S H
Division of Food Science & Biotechnology, College of Agriculture & Life Sciences, Kangwon National University, Chunchon, Korea.
Mol Cells. 2001 Feb 28;11(1):7-12.
With the long-term goal of generating CMV-resistant transgenic plants using antibody genes, a single-chain variable fragment (scFv) antibody that binds to the cucumber mosaic virus was isolated from a scFv phage display library by four rounds of affinity selection with CMV-Mf as an antigen. The scFv has the identical binding specificity to CMV as a monoclonal antibody that is generated by the hybridoma fusion technique, and recognized purified preparations of CMV isolates belonging to either subgroup I or II in immunoblotting. The nucleotide sequences of the recombinant antibody showed that a heavy chain variable region (V(H)) gene belonged to the VH3 subgroup and the kappa light chain variable region (V kappa) came from the Vkappa4 subgroup. Our results demonstrate that the scFv phage display library, an alternative approach to the traditional hybridoma fusion technique, has a potential applicability in the study of plant virus and plant pathology.
以利用抗体基因培育抗黄瓜花叶病毒（CMV）转基因植物为长期目标，通过以CMV-Mf为抗原进行四轮亲和筛选，从单链可变片段（scFv）噬菌体展示文库中分离出一种与黄瓜花叶病毒结合的scFv抗体。该scFv与通过杂交瘤融合技术产生的单克隆抗体对CMV具有相同的结合特异性，并且在免疫印迹中识别属于I或II亚组的CMV分离株的纯化制剂。重组抗体的核苷酸序列表明，重链可变区（V(H)）基因属于VH3亚组，κ轻链可变区（Vκ）来自Vκ4亚组。我们的结果表明，scFv噬菌体展示文库作为传统杂交瘤融合技术的替代方法，在植物病毒和植物病理学研究中具有潜在的适用性。
