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Abstract：
Avian leukosis virus (ALV) is an avian oncogenic retrovirus that can impair immunological function, stunt growth and decrease egg production in avian flocks. The capsid protein (P27) is an attractive candidate for ALV diagnostics. In the present study, a new hybridoma cell (1F8) stably secreting an anti-P27 monoclonal antibody (mAb) was developed. The mAb exhibited a high affinity constant (Ka) of 8.65 × 106.0 L/mol, and it could be used for the detection of ALV-A/B/J/K strains. Moreover, a total of eight truncated recombinant proteins and five synthetic polypeptides were utilized for the identification of the B-cell epitopes present on P27. The results revealed that 218IIKYVLDRQK227 was the minimal epitope recognized by 1F8, which had never been reported before. Additionally, the epitopes could strongly react with different ALV subgroup\'s specific positive serum and had a complete homology among all the ALV subgroups strains. Finally, a new sandwich ELISA method was created for the detection of ALV antigens, demonstrating increased sensitivity compared to a commercially available ELISA kit. These results offer essential knowledge for further characterizing the antigenic composition of ALV P27 and will facilitate the development of diagnostic reagents for ALV.
摘要：
禽白血病病毒(ALV)是一种可损害免疫功能的禽致癌逆转录病毒,抑制禽群的生长和减少产蛋量。衣壳蛋白(P27)是ALV诊断的有吸引力的候选者。在本研究中,开发了稳定分泌抗P27单克隆抗体（mAb）的新杂交瘤细胞（1F8）。mAb具有8.65×106.0L/mol的高亲和力常数(Ka),可用于ALV-A/B/J/K菌株的检测。此外，使用总共八种截短的重组蛋白和五种合成多肽来鉴定P27上存在的B细胞表位。结果表明，218IIKYVLDRQK227是1F8识别的最小表位，以前从未报道过。此外,这些表位能与不同ALV亚组的特异性阳性血清发生强烈反应，并且在所有ALV亚组菌株之间具有完全同源性。最后,建立了一种新的夹心ELISA方法来检测ALV抗原，与市售ELISA试剂盒相比，显示出更高的灵敏度。这些结果为进一步表征ALVP27的抗原组成提供了必要的知识，并将促进ALV诊断试剂的开发。
