Meqdam M, Mostratos A
Department of Cell and Structural Biology, Medical School, University of Manchester, UK.
New Microbiol. 1999 Jan;22(1):19-25.
A computer program combining of hydrophilicity, flexibility, surface probability, secondary structure and antigenic index parameters of the amino acid sequence of measles virus (MV) fusion protein was used to select four possible epitopes. Rabbits were immunized with the synthesized peptides conjugated to purified protein derivative using the homobifunctional cross-linker bis-sulfosuccinimidyl suberate. Immune stimulating complexes were prepared with the peptides conjugated to the purified protein derivative carrier using a dialysis method. All antisera raised in rabbits against the peptide conjugates had a high titer to the homologous peptides and reacted well with denatured MV as tested by plate ELISA. None of the sera had neutralizing antibody. Human sera positive for MV antibody reacted strongly with the synthesized peptides indicating that the selected locations function as partial antigenic sites. Antisera against peptide conjugates reacted weakly in immunofluorescence and none of these antisera reacted with purified MV proteins in Western blot. The results obtained in this study indicated that although the computer program could not predict epitopes important for the neutralization of the MV, the predicted epitopes are useful for detecting antibodies against MV.
使用一个结合了麻疹病毒（MV）融合蛋白氨基酸序列的亲水性、柔韧性、表面可能性、二级结构和抗原指数参数的计算机程序来选择四个可能的表位。使用同双功能交联剂双琥珀酰亚胺基辛二酸酯将合成肽与纯化蛋白衍生物偶联，对兔子进行免疫。采用透析法用与纯化蛋白衍生物载体偶联的肽制备免疫刺激复合物。通过平板ELISA检测，所有在兔子体内产生的针对肽偶联物的抗血清对同源肽具有高滴度，并且与变性的MV反应良好。没有一种血清具有中和抗体。对MV抗体呈阳性的人血清与合成肽强烈反应，表明所选位置起部分抗原位点的作用。针对肽偶联物的抗血清在免疫荧光中反应较弱，并且这些抗血清在蛋白质印迹中均未与纯化的MV蛋白反应。本研究获得的结果表明，尽管计算机程序无法预测对MV中和重要的表位，但预测的表位可用于检测抗MV抗体。
