Pauillac S, Halmos T, Labrousse H, Antonakis K, Avrameas S
CNRS URA 359, Département d'Immunologie, Institut Pasteur, Paris, France.
J Immunol Methods. 1993 Sep 15;164(2):165-73. doi: 10.1016/0022-1759(93)90309-u.
Monensin, a polyether antibiotic of molecular weight 671 Da, was converted into a hemisuccinate and covalently linked to bovine serum albumin via the mixed anhydride method. Using this immunogen, polyclonal anti-monensin antibodies were raised in rabbits and monoclonal antibodies were prepared from mice. The specificity of the anti-monensin antibodies was examined by using several structural analogues as the immunogen and by performing direct binding and competitive microELISA assays on Terasaki plates. Rabbit polyclonal antibodies had a dissociation constant (KD) of 5.5 x 10(-8) M for monensin and reacted with nigericin, an antibiotic structurally related to monensin. In contrast, a mouse monoclonal antibody, 2H8, reacted only with monensin and had a much lower KD = 3 x 10(-8) M for monensin. Monoclonal antibody 2H8 was used to develop a competitive microELISA able to detect as little as 5 ng/ml of monensin in solution which corresponds to 75 pg or 110 fmol of this hapten per Terasaki well.
莫能菌素是一种分子量为671道尔顿的聚醚抗生素，它被转化为半琥珀酸酯，并通过混合酸酐法与牛血清白蛋白共价连接。使用这种免疫原，在兔子体内产生了多克隆抗莫能菌素抗体，并从小鼠体内制备了单克隆抗体。通过使用几种结构类似物作为免疫原，并在Terasaki板上进行直接结合和竞争性微酶联免疫吸附测定，对抗莫能菌素抗体的特异性进行了检测。兔多克隆抗体对莫能菌素的解离常数（KD）为5.5×10⁻⁸ M，并与尼日利亚菌素发生反应，尼日利亚菌素是一种与莫能菌素结构相关的抗生素。相比之下，小鼠单克隆抗体2H8仅与莫能菌素发生反应，对莫能菌素的KD值低得多，为3×10⁻⁸ M。单克隆抗体2H8被用于开发一种竞争性微酶联免疫吸附测定法，该方法能够检测溶液中低至5 ng/ml的莫能菌素，这相当于每个Terasaki孔中有75 pg或110 fmol的这种半抗原。
