Antibodies with infinite affinity
Antibodies with infinite affinity
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DOI：
10.1073/pnas.151260298
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发表时间：
2001-07-17
影响因子：
11.1
通讯作者：
Meares, CF
中科院分区：
文献类型：
--
作者：
Chmura, AJ;Orton, MS;Meares, CF
在这里，我们报告的方法来设计和生产的抗体/配体对，以实现远大于亲和素/生物素的功能亲和力。使用基本的化学原理，我们已经开发了保留抗体的结合特异性但不解离的抗体/配体对。选择结构表征的抗体/配体对作为示例，我们设计了抗体结合口袋和配体中的互补反应基团，使得它们在抗体/配体复合物中紧密接近。由于这些基团的低反应性，避免了与介质中其他分子的交叉反应;然而，在抗体/配体复合物中，互补反应基团的有效局部浓度非常大，允许共价反应将两者连接在一起。通过消除配体与抗体的解离，我们使亲和力在功能上无限大。这种亲和力的化学操作适用于其他生物结合对。
Here we report an approach to the design and production of antibody/ligand pairs, to achieve functional affinity far greater than avidin/biotin. Using fundamental chemical principles, we have developed antibody/ligand pairs that retain the binding specificity of the antibody, but do not dissociate. Choosing a structurally characterized antibody/ligand pair as an example, we engineered complementary reactive groups in the antibody binding pocket and the ligand, so that they would be in close proximity in the antibody/ligand complex. Cross-reactions with other molecules in the medium are averted because of the low reactivity of these groups; however, in the antibody/ligand complex the effective local concentrations of the complementary reactive groups are very large, allowing a covalent reaction to link the two together. By eliminating the dissociation of the ligand from the antibody, we have made the affinity functionally infinite. This chemical manipulation of affinity is applicable to other biological binding pairs.