Freeman J W, Chatterjee A, Busch H
J Immunol Methods. 1985 Apr 22;78(2):259-65. doi: 10.1016/0022-1759(85)90083-3.
A new method is described for determining the distribution of epitopes identified by monoclonal antibodies. The method utilizes nitrocellulose membranes as a solid support for antigens which are rapidly adsorbed to nitrocellulose by vacuum-blotting and then used in competitive antibody binding assays. The distribution of epitopes is established by the reciprocal cross-blocking of radiolabeled antibody by increasing concentrations of unlabeled antibody. When unlabeled antibody does not block the binding of labeled antibody to antigen, the 2 antibodies recognize distinct epitopes. When unlabeled antibody blocks the binding of labeled antibody to antigen, the 2 antibodies recognize the same epitope. The method is rapid, sensitive and should be applicable to screening monoclonal antibodies to any epitope.
本文描述了一种用于确定单克隆抗体识别的表位分布的新方法。该方法利用硝酸纤维素膜作为抗原的固相支持物，抗原通过真空印迹快速吸附到硝酸纤维素上，然后用于竞争性抗体结合试验。通过增加未标记抗体的浓度，用放射性标记抗体进行相互交叉阻断来确定表位的分布。当未标记抗体不阻断标记抗体与抗原的结合时，这两种抗体识别不同的表位。当未标记抗体阻断标记抗体与抗原的结合时，这两种抗体识别相同的表位。该方法快速、灵敏，应适用于筛选针对任何表位的单克隆抗体。
