Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, MD, 20850, USA.
Centre for Oncology, Radiopharmaceuticals and Research, Biologics and Radiotherapeutic Drugs Directorate, Health Canada, 251 Sir Frederick Banting Driveway, K1A 0K9, Ottawa, ON, Canada.
Biomol NMR Assign. 2023 Jun;17(1):75-81. doi: 10.1007/s12104-023-10123-9. Epub 2023 Mar 1.
The monoclonal antibody (mAb) protein class has become a primary therapeutic platform for the production of new life saving drug products. MAbs are comprised of two domains: the antigen-binding fragment (Fab) and crystallizable fragment (Fc). Despite the success in the clinic, NMR assignments of the complete Fab domain have been elusive, in part due to problems in production of properly folded, triply-labeled H,C,N Fab domain. Here, we report the successful recombinant expression of a triply-labeled Fab domain, derived from the standard IgG1κ known as NISTmAb, in yeast. Using the H,C,N Fab domain, we assigned 94% of the H, C, and N backbone atoms.
单克隆抗体 (mAb) 蛋白类已成为生产新的救命药物产品的主要治疗平台。单克隆抗体由两个结构域组成：抗原结合片段 (Fab) 和可结晶片段 (Fc)。尽管在临床上取得了成功，但完整 Fab 结构域的 NMR 分配仍然难以捉摸，部分原因是生产适当折叠的三重标记 H、C、N Fab 结构域存在问题。在这里，我们报告了在酵母中成功表达了一种三重标记的 Fab 结构域，该结构域源自标准 IgG1κ，称为 NISTmAb。使用 H、C、N Fab 结构域，我们分配了 94%的 H、C 和 N 骨架原子。
