摘要
目的 分析汉滩病毒核衣壳蛋白 (NP)主要线性表位免疫学反应的最小抗原多肽的组成 .方法 在应用多株国产单克隆抗体和噬菌体文库技术基本确定了病毒 NP氨基端的一个主要的 B细胞线性抗原表位的基础上 ,进一步用单克隆抗体 L 13F3对 6肽或 15肽随机多肽文库进行筛选 ,并根据病毒 NP氨基端 aa15 - 6 7的氨基酸序列 ,在纤维素膜上每间隔 1或 2个氨基酸合成了 15肽和 8肽的阵列 ,用 L13F3对这两组多肽阵列进行免疫学检测 .结果 对随机多肽文库进行筛检未获阳性结果 ,而对两组多肽阵列的检测确定了 2个分别由 5肽和 3肽组成的最小的抗原反应多肽 .结论 多肽阵列检测技术即肽扫描 (peptide scan)
AIM To map the minimal epitopes of Hantaan virus nucleocapsid protein with monoclonal antibody (MaAb) L13F3. METHODS After our biopanning with some strains of MaAbs and the phage library of Hantaan virus S gene-peptide fragments and sequencing for positive clonies, the bio-panning and screening of 6mer and 15mer random peptide libraries was performed, and two groups of peptides arraies synthesized following the sequence of aa15-67 of ORF of Hantaan virus nucleocapsid protein were scanned with MaAb L13F3. RESULTS MaAb L13F3 did not react with any of random peptide libraries. Peptides scanning revealed that it required minimal epitopes of 5 or 3 amino acids for antigen recognition. CONCLUSION The pepscan is an exact, effective and convenient method for fine mapping of epitopes.
出处
《第四军医大学学报》
2000年第7期850-852,共3页
Journal of the Fourth Military Medical University
基金
中国-欧共体合作科研基金!资助 [CI1* -CT94-0 0 2 4( DG 12HSMU) ]
