Backbone NMR assignment of the yeast expressed Fab fragment of the NISTmAb reference antibody

Abstract

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 ²H,¹³C,¹⁵N 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 ²H,¹³C,¹⁵N Fab domain, we assigned 94% of the ¹H, ¹³C, and ¹⁵N backbone atoms.