Department of Molecular and Medical Pharmacology, University of California, Los Angeles, USA.
Angew Chem Int Ed Engl. 2011 Aug 29;50(36):8295-8. doi: 10.1002/anie.201101149. Epub 2011 Jul 14.
Since the invention of hybridoma technology, methods for generating affinity reagents that bind specific target molecules have revolutionized biology and medicine. In the postgenomic era, there is a pressing need to accelerate the pace of ligand discovery to elucidate the functions of a rapidly growing number of newly characterized molecules and their modified states. Nonimmunoglobulin-based proteins such as DARPins, affibodies, and monobodies represent attractive alternatives to traditional antibodies as these are small, soluble, disulfide-free, single-domain scaffolds that can be selected from combinatorial libraries and expressed in bacteria. For example, monobodies—highly stable scaffolds based on the immunoglobulin VH-like 10th fibronectin type III (10Fn3) domain of human fibronectin—have yielded antibody mimetics that bind to numerous targets for applications including intracellular inhibition, therapeutics, and biosensors. These 10Fn3-based ligands can be derived from highly diverse libraries using techniques such as phage, ribosome, mRNA, bacterial, and yeast displays.
自杂交瘤技术发明以来，生成与特定靶分子结合的亲和试剂的方法彻底改变了生物学和医学。在后基因组时代，迫切需要加快配体发现的步伐，以阐明大量新表征的分子及其修饰状态的功能。像 DARPins、affibodies 和 monobodies 这样的非免疫球蛋白蛋白是传统抗体的有吸引力的替代品，因为它们是小的、可溶性的、无二硫键的、单结构域支架，可以从组合文库中选择并在细菌中表达。例如，monobodies——基于人纤连蛋白的免疫球蛋白 VH 样第 10 个纤维连接蛋白 III 型（10Fn3）结构域的高度稳定的支架——产生了与许多靶标结合的抗体模拟物，可用于包括细胞内抑制、治疗和生物传感器在内的应用。这些基于 10Fn3 的配体可以使用噬菌体、核糖体、mRNA、细菌和酵母展示等技术从高度多样化的文库中衍生出来。
