Picha Kristen, Huang Chichi, Bugelski Peter, O'Neil Karyn
Centocor Research and Development, Inc., A Division of Johnson & Johnson Company, Radnor, PA, USA.
Methods Mol Biol. 2014;1088:125-45. doi: 10.1007/978-1-62703-673-3_9.
The MIMETIBODY™ platform was developed to expand the opportunities for application of biotherapeutics. While the utility of antibodies as antagonists has been well demonstrated, their application as agonists has been more challenging. For steric reasons, antibodies may be less well suited to perform as agonists or as inhibitors of GPCRs. In contrast, many bioactive peptides function as agonists or by interaction with GPCRs but their development as therapeutics has been challenging due to their small size and metabolic lability. The MIMETIBODY™ platform has been used to develop a variety of stable, long-lived molecules with intrinsic activities similar to that of their parent peptides. This chapter describes methods for construction of expression plasmids, expression and purification strategies, and methods for characterizing the activity of these novel proteins.
MIMETIBODY™平台的开发旨在拓展生物治疗药物的应用机会。虽然抗体作为拮抗剂的效用已得到充分证明，但其作为激动剂的应用则更具挑战性。由于空间位阻原因，抗体可能不太适合作为G蛋白偶联受体（GPCR）的激动剂或抑制剂。相比之下，许多生物活性肽作为激动剂发挥作用或通过与GPCR相互作用发挥作用，但其作为治疗药物的开发一直具有挑战性，因为它们体积小且代谢不稳定。MIMETIBODY™平台已被用于开发各种具有与母体肽相似内在活性的稳定、长效分子。本章描述了表达质粒构建方法、表达和纯化策略以及这些新型蛋白质活性表征方法。
