Ochoa M, Bárcena J, de la Luna S, Melero J A, Douglas A R, Nieto A, Ortín J, Skehel J J, Portela A
Instituto de Salud Carlos III, Centro Nacional de Biología Celular y Retrovirus, Madrid, Spain.
Virus Res. 1995 Aug;37(3):305-15. doi: 10.1016/0168-1702(95)00039-s.
Characterization of the epitopes recognized by 21 monoclonal antibodies (MAbs) specific for the influenza A virus PA (13 MAbs) and PB2 (8 MAbs) polypeptides (Bárcena et al. (1994) J. Virol. 68, 6900-6909) raised against denatured polypeptides produced in E. coli is described. MAbs were characterized by: (1) competitive binding ELISAs; (2) mapping of the protein regions that specify their binding sites; and (3) analyses of their ability to recognize the corresponding viral protein in a number of viral isolates. Five and three non-overlapping antigenic areas were defined by the anti-PA and anti-PB2 MAbs, respectively. Five of the anti-PA MAbs recognized antigenic determinants located within the amino-terminal 157 amino acids of the PA protein, and 6 others reacted strongly with a PA fragment comprising the first 236 amino acids. All 8 anti-PB2 antibodies reacted strongly with a polypeptide fragment containing amino acids 1-113 of the PB2 protein. Analyses of the reactivities of 4 anti-P antibodies with 23 influenza A virus reference strains isolated over a period of 61 years and recovered from humans, pigs, birds and horses, showed that the epitopes were conserved among all viral isolates. The application of these antibodies as research and diagnostic tools is discussed.
本文描述了针对甲型流感病毒PA（13种单克隆抗体）和PB2（8种单克隆抗体）多肽产生的21种单克隆抗体（MAb）所识别表位的特性（Bárcena等人，（1994年）《病毒学杂志》68卷，6900 - 6909页），这些单克隆抗体是针对在大肠杆菌中产生的变性多肽制备的。通过以下方法对单克隆抗体进行了特性鉴定：（1）竞争性结合酶联免疫吸附测定（ELISA）；（2）确定其结合位点的蛋白质区域图谱绘制；（3）分析它们在多种病毒分离株中识别相应病毒蛋白的能力。抗PA和抗PB2单克隆抗体分别确定了5个和3个不重叠的抗原区域。5种抗PA单克隆抗体识别位于PA蛋白氨基末端157个氨基酸内的抗原决定簇，另外6种与包含前236个氨基酸的PA片段强烈反应。所有8种抗PB2抗体均与包含PB2蛋白第1 - 113位氨基酸的多肽片段强烈反应。对4种抗P抗体与61年间从人、猪、鸟和马中分离得到的23种甲型流感病毒参考毒株反应性的分析表明，所有病毒分离株中的表位都是保守的。本文还讨论了这些抗体作为研究和诊断工具的应用。
