Mapping the binding of monoclonal antibodies to the acetylcholine receptor from Torpedo californica.
Mapping the binding of monoclonal antibodies to the acetylcholine receptor from Torpedo californica.
复制标题
绘制单克隆抗体与加州鱼雷乙酰胆碱受体的结合图谱。
DOI：
10.1021/bi00283a003
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发表时间：
1983
期刊：
影响因子：
2.9
通讯作者：
Lindstrom,JM
中科院分区：
文献类型：
--
作者：
Gullick,WJ;Lindstrom,JM
摘要：对来自几个物种的154株抗乙酰胆碱受体的单抗(MAbs)进行了结合能力的检测。检测到74株mAb结合，免疫沉淀和凝胶电泳法确定了它们的主要亚基结合偏好。几种单抗在亚基之间发生交叉反应。为了定量它们的相对亲和力，我们测定了每一单抗针对每一亚单位的表观效价。94%的单抗对单个亚基至少有95%的特异性。大多数交叉反应的单抗主要与一个亚基结合，与其他亚基结合较弱。然而，两个单抗在一定程度上与所有四个亚基结合。为了评估它们结合的特异性，我们测试了它们与牛血清白蛋白、卵清蛋白和磷高磷蛋白的结合。只有一株单抗与磷高磷蛋白发生了外来的交叉反应。我们还测试了5株抗无关抗原的单抗，这些单抗具有潜在的潜在优点。单抗作为乙酰胆碱受体的结构和功能的探针，是可以针对受体大分子的多个部分制备的。原则上，mAb结合的位置可以相当准确地确定。然而，将单抗的文库映射到受体并由此表征受体的抗原结构的第一步是简单地确定每一单抗的受体亚单位特异性，然后确定在每一受体亚单位内可以检测到多少不同的抗原决定簇。在这篇论文中，我们利用来自Torpedo calfornica的受体的亚单位，为154个mAb库进行了这些第一步，该库针对来自Torpedo calfornica和Electric Electricus以及牛和人的肌肉的受体(Tzartos&Lindstrom，1980,1981；Tzartos等人，1981；SJ Tzartos，LW Swanson，L.K.Langeberg，S.M.Hochschwender和J.M.Lindstrom，未发表的结果)。在随后的论文中，我们将沿着a(W.J.Gullick等人，未发表的结果)和8个亚基(Anderson等人，1983)的氨基酸序列对这些抗原决定簇和亚基的其他结构特征进行排序。来自加利福尼亚鱼雷和电鱼电子器官的乙酰胆碱受体具有亚单位化学计量比a2py8(Reynolds&Karlin，1978；Lindstrom等人，1979a；Raftery等人，1980；Conti-Tronconi等人，1982b)。这表明加州圣地亚哥索尔克生物研究所的f亚基化学计量比
W. J. Gullick** and J. M. Lindstrom* abstract: One hundred and fifty-four monoclonal antibodies (mAbs) raised against acetylcholine receptors from several species were tested for their ability to bind iodinated dissociated receptor subunits from Torpedo californica electric organ. Seventy-four mAbs bound detectably, and their primary subunit binding preference was established by immunopre-cipitation followed by gel electrophoresis. Several mAbs cross-reacted between subunits. In order to quantitate their relative affinity we determined the apparent titer of each mAb against each subunit. Ninety-four percent of the mAbs tested were at least 95% specific for a single subunit. Most cross-reacting mAbs bound predominantly to one subunit and weakly to others. However, two mAbs bound to some degree with all four subunits. To assess their specificity of binding, we tested them all for binding to bovine serum albumin, ovalbumin, and phosvitin. Only one mAb which bound to phosvitin showed any extraneous cross-reaction. We also tested five mAbs raised against unrelated antigens for reaction with great potential virtueof monoclonal antibodies (mAbs) as probes of the structure and function of acetylcholine re-ceptors is that mAbs can be prepared against many parts of the receptor macromolecule. In principal, the site at which a mAb binds can be determined rather precisely. However, the first steps in mapping a library of mAbs to the receptor, and thereby characterizing the antigenic structure of the re-ceptor, are simply to determine the receptor subunit specificity of each mAb and then determine how many different antigenic determinants can be detected within each receptor subunit. In this paper we undertake these first steps for a library of 154 mAbs raised against receptors from the electric organs of Torpedo californica and Electrophorus electricus and the muscles of cattle and humans (Tzartos & Lindstrom, 1980, 1981; Tzartos et al., 1981; SJ Tzartos, LW Swanson, L. K. Langeberg, S. M. Hochschwender, and J. M. Lindstrom, unpublished results) using subunits of receptor from Torpedo californica. In subsequent papers we will order these antigenic determinants and other structural features of the subunits along the amino acid sequence of the a (W. J. Gullick et al., unpublished results) and 8 subunits (Anderson et al., 1983). Acetylcholine receptors from the electricorgans of Torpedo californica and Electrophorus electricus have the subunit stoichiometry a2py8 (Reynolds & Karlin, 1978; Lindstrom et al., 1979a; Raftery et al., 1980; Conti-Tronconi et al., 1982b). This indicates that the subunit stoichiometry of f From the Salk Institute for Biological Studies, San Diego, California