Study on production and utilization of plant virus specific antibodies using anti-synthetic peptide antibodies

研究结果摘要;三叶草黄脉病毒（CYVV）和菜豆黄花叶病毒（BYMV）是两种生物学和血清学性质非常接近的马铃薯Y病毒属病毒，用多克隆抗体和单克隆抗体很难区分它们。根据外壳蛋白的氨基酸序列，利用SPOTs系统进行抗原表位分析，在外壳蛋白N端检测到CYVV-NFU的5个抗原表位，其中一个抗原表位是BYMV和CYVV共有的抗原表位，利用合成的多肽制备了抗肽抗体。该抗体用于ELISA、Western blot、ISEM等血清学方法检测CYVV和BYMV均无非特异性反应，SPOTs系统也可用于单克隆抗体的抗原表位检测，但部分抗原表位的抗血清效价较低，未能成功制备。结果表明，SPOTs检测的表位所获得的肽段并不总是可以作为强抗原，或者某些抗肽抗体不适合用于某些血清学诊断。

SUMMARY OF RESEARCH RESULTS ; Two potyviruses, clover yellow vein virus (CYVV) and bean yellow mosaic virus (BYMV) are biologically and serologically very colse and it is difficult to distinguish them by the use of poly- and monoclonal antibodies. Upon the information of amino acid sequence of coat protein, the epitope analysis by SPOTs system was conducted and 5 epitopes in N terminals of coat protein region for CYVV-NFU were detected.As one epitope was common in both BYMV and CYVV, anti-peptide antibody was produced using synthetic peptides. The antibody was used in ELISA, Western blot, ISEM and other serological method to detect CYVV and BYMV without non specific reaction.SPOTs system was also shown to be useful to detect epitopes which are reacted with monoclonal antibodies in the experiment using monoclonal antibody obtained from ATCC.Production of some anti-peptide antibodies to some epitopes, however, was not successful because only the antisera with low titer were obtained. It showed that peptides obtained from epitopes detected by SPOTs can not always be used as strong antigens or some anti-peptide antibodies are not suitable to use in some serodiagnosis.