Shukla D D, Tribbick G, Mason T J, Hewish D R, Geysen H M, Ward C W
Commonwealth Scientific and Industrial Research Organization, Division of Biotechnology, Parkville Laboratory, Australia.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8192-6. doi: 10.1073/pnas.86.21.8192.
Virus-specific or group-specific antibody probes to potyviruses can be produced by targeting the immune response to the virus-specific, N-terminal region of the capsid protein (29-95 amino acids depending on the virus) or to the conserved core region (216 amino acids) of the capsid protein, respectively. Immunochemical analysis of overlapping, synthetic octapeptides covering the capsid protein of the Johnsongrass strain of Johnsongrass mosaic virus (JGMV-JG) has delineated the peptide sequences recognized by five polyclonal rabbit antisera and two mouse monoclonal antibodies (mAbs). The antibodies characterized were (i) three virus-specific rabbit polyclonal antisera and one virus-specific mouse mAb (1/25) raised against native virus particles, (ii) one polyclonal antiserum raised against trypsin-derived core particles of JGMV-JG, (iii) one group-specific polyclonal antiserum raised against the denatured, truncated coat protein from trypsin-derived core particles of JGMV-JG, and (iv) one group-specific mouse mAb (1/16) raised against native virus particles. The two epitopes seen by mAb 1/25 occurred at residues 18-27 and 43-52 and overlapped with the two major epitopes seen by the virus-specific polyclonal antiserum. The group-specific epitope seen in JGMV-JG by mAb 1/16 was also recognized strongly in potato virus Y, the type member of the potyvirus group. The multiple epitopes seen by the cross-reactive polyclonal antisera were distributed across the entire core region of the coat protein and their relative antibody binding responses varied between JGMV-JG, potato virus Y, and six other distinct potyviruses.
针对马铃薯Y病毒属病毒的病毒特异性或组特异性抗体探针可以通过分别针对衣壳蛋白病毒特异性的N端区域（根据病毒不同，含29 - 95个氨基酸）或衣壳蛋白的保守核心区域（216个氨基酸）引发免疫反应来制备。对覆盖高粱花叶病毒约翰逊草株系（JGMV-JG）衣壳蛋白的重叠合成八肽进行免疫化学分析，已确定了五种兔多克隆抗血清和两种小鼠单克隆抗体（mAb）所识别的肽序列。所鉴定的抗体包括：（i）三种针对天然病毒颗粒产生的病毒特异性兔多克隆抗血清和一种病毒特异性小鼠单克隆抗体（1/25）；（ii）一种针对JGMV-JG胰蛋白酶消化产生的核心颗粒产生的多克隆抗血清；（iii）一种针对JGMV-JG胰蛋白酶消化产生的核心颗粒变性、截短的外壳蛋白产生的组特异性多克隆抗血清；（iv）一种针对天然病毒颗粒产生的组特异性小鼠单克隆抗体（1/16）。单克隆抗体1/25识别的两个表位位于第18 - 27位氨基酸残基和第43 - 52位氨基酸残基，与病毒特异性多克隆抗血清识别的两个主要表位重叠。单克隆抗体1/16在JGMV-JG中识别的组特异性表位在马铃薯Y病毒（马铃薯Y病毒属的模式种）中也能被强烈识别。交叉反应多克隆抗血清识别的多个表位分布在外壳蛋白的整个核心区域，它们在JGMV-JG、马铃薯Y病毒和其他六种不同的马铃薯Y病毒属病毒之间的相对抗体结合反应有所不同。
