Friguet B, Fedorov A, Serganov A A, Navon A, Goldberg M E
Unité de Biochimie Cellulaire (CNRS URA 1129), Institut Pasteur, Paris, France.
Anal Biochem. 1993 May 1;210(2):344-50. doi: 10.1006/abio.1993.1206.
This communication describes a novel highly sensitive method for measuring the affinity of a monoclonal antibody for its antigen. It is based on a radioimmunoassay in which the antigen is labeled with radioactivity. It is therefore particularly well adapted to the study of trace amounts of radiolabeled polypeptide chains produced either in vivo, or in vitro by a cell free protein synthesis system or by chemical radiolabeling. It offers several advantages over previously described methods. Though making use of insolubilized antibody, it does measure the true affinity constant of the monoclonal antibody in solution for the antigen. It can be used even when the antigen is present at concentrations far below the dissociation constant of the antibody/antigen complex. It does not require the antigen or the antibody to be purified. In most cases, it requires no sophisticated equipment. This method could be easily adapted to the determination of the equilibrium constant of any type of protein/ligand system.
本通讯描述了一种用于测量单克隆抗体与其抗原亲和力的新型高灵敏度方法。它基于一种放射免疫测定法，其中抗原用放射性标记。因此，它特别适合于研究体内或体外由无细胞蛋白质合成系统或化学放射性标记产生的痕量放射性标记多肽链。与先前描述的方法相比，它具有几个优点。尽管使用了不溶性抗体，但它确实能测量溶液中单克隆抗体对抗原的真实亲和力常数。即使抗原浓度远低于抗体/抗原复合物的解离常数，也可以使用该方法。它不需要纯化抗原或抗体。在大多数情况下，它不需要复杂的设备。这种方法可以很容易地适用于任何类型蛋白质/配体系统平衡常数的测定。
