Codon optimization regulates IgG3 and IgM expression and glycosylation in N. benthamiana

• DOI: 10.3389/fbioe.2023.1320586
• Publication Date: 2023-12-06T10:33:16Z
• Authors: Lin Sun; Somanath Kallolimath; Roman Palt; Florian Eminger; Richard Strasser; Herta Steinkellner
• Keywords: 0303 health sciences; 03 medical and health sciences; codon usage; glycosylation; antibody; Nicotiana benthamiana; Bioengineering and Biotechnology; transcription and translation; TP248.13-248.65; Biotechnology

Plants are being increasingly recognized for the production of complex human proteins, including monoclonal antibodies (mAbs). Various methods have been applied to boost recombinant expression, with DNA codon usage an important approach. Here, we transiently expressed three mAbs in Nicotiana benthamiana, namely one IgG3 and two IgM directed against SARS-CoV-2 as optimized(CO) non-codon optimized (NCO) variants. qRT-PCR exhibited significantly increased mRNA levels all CO variants compared orthologues, line protein expression. Purified NCO did not exhibit obvious biochemical differences, determined by SDS-PAGE antigen binding activities. By contrast, enhanced selectively impacts on glycosite occupancy N-glycan processing, mannosidic structures. The results point a careful monitoring proteins upon enhancing Especially if it comes therapeutic application even subtle modifications might alter product efficacy or increase immunogenicity.