Rapid Production of Cyclic Citrullinated Peptide Monoclonal Antibody in Nicotiana benthamiana for the Early Detection and Diagnosis of Rheumatoid Arthritis

Abstract

Rheumatoid arthritis (RA) is one of the most common autoimmune diseases, affecting 0.5% to 1% of the population. It could ultimately result in joint destruction, functional decline, work disability, and enhanced mortality. Cyclic citrullinated peptide antibodies (CCP Abs) are useful biomarkers for the early detection and diagnosis of RA. In this study, we used plant viral-based expression vectors that produce rapidly large quantities of CCP-specific monoclonal antibodies. Heavy and light chain genes of a CCP monoclonal antibody (CCP mAb) were cloned from the hybridoma cell (12G1) and introduced into two separate plant viral-based expression vectors, TMV and PVX. A cyclic citrullinated peptide monoclonal antibody was produced in Nicotiana benthamiana through an Agrobacterium-mediated transient expression system. The expression of CCP mAb in tobacco plants was confirmed by dot blot, western blot analysis, and enzyme-linked immunosorbent assays (ELISA). It was shown that tobacco plants could accumulate CCP mAbs up to 0.35% of total soluble protein. Accumulated CCP mAb from infiltrated leaves was purified by protein G affinity chromatography. Immunoblot assays and ELISA showed plant-produced CCP mAbs successfully bound to a synthetic CCP peptide antigen. This system provides a fast strategy for the production of pharmaceutical CCP mAbs in tobacco plants.

The Synergistic Effects of Environmental and Genetic Factors on the Regulation of Anthocyanin Accumulation in Plant Tissues

“…This vector expression system has the advantage of rapid replication of insert genes because a part of them is constructed by recombining from viral mRNA replicon [ 39 , 40 ]. Using this vector expression system allows rapid and high-yield production of recombinant protein in plants [ 41 , 42 ]. The fruits infiltrated with anthocyanin biosynthesis genes were much more colorful than the control (MOCK) fruits ( Figure 2 and Figure 3 ).…”

SynBio: A Journal for Advancing Solutions to Global Challenges

“…In the Special Issue "Feature Papers in Synthetic Biology", Do [11] focused on rheumatoid arthritis, a common autoimmune disease that can cause joint destruction and disability (1b and 2a). In this study, Do used plant virus-based expression vectors to produce large quantities of CCP-specific monoclonal antibodies (mAbs).…”