Abstract
Delivery of secretory immunoglobulin A (sIgA) to mucosal surfaces as a passive immunotherapy agent is a promising strategy to prevent infectious diseases. Recombinant sIgA production in plants requires the co-expression of four transcriptional units encoding the light chain (LC), heavy chain (HC), joining chain (JC) and secretory component (SC). As a way to optimize sIgA production in plants, we tested the combinatorial expression of 16 versions of a human sIgA against the VP8* rotavirus antigen in Nicotiana benthamiana, using the recently developed GoldenBraid multigene assembly system. Each sIgA version was obtained by combining one of the two types of HC (α1 and α2) with one of the two LC types (k and λ) and linking or not a KDEL peptide to the HC and/or SC. From the analysis of the anti-VP8* activity, it was concluded that those sIgA versions carrying HCα1 and LCλ provided the highest yields. Moreover, ER retention significantly increased antibody production, particularly when the KDEL signal was linked to the SC. Maximum expression levels of 32.5 μg IgA/g fresh weight (FW) were obtained in the best performing combination, with an estimated 33% of it in the form of a secretory complex.
Highlights
Monoclonal antibodies have been used in research and diagnosis for many years, and their application in health is increasing rapidly
IgA can be present in the body fluids in its monomeric form, containing only heavy chain (HC) and light chain (LC) or forming a secretory IgA, a multiprotein structure comprising two full IgA molecules dimerized by a short joining chain (JC) and surrounded by the secretory component (SC), a polypeptide resulting from the proteolytic cleavage of the poly-immunoglobulin receptor
There are a number of options available when designing a new antibody in secretory IgA (sIgA) format, which could lead to completely different products for the same purpose
Summary
Monoclonal antibodies (mAbs) have been used in research and diagnosis for many years, and their application in health is increasing rapidly. There are a number of options available when designing a new antibody in sIgA format, which could lead to completely different products for the same purpose. MoClo [29], to achieve all the combinations should be facilitated These technologies open a new way for optimization of antibody production in plants by experimentally testing the best subcellular targeting and isotype combinations for the expression of a target antibody. The variable regions of 2A1 antibody were initially selected by phage display against the VP8* peptide of the VP4 protein of the rotavirus SA11 strain capsid [30] This antibody was previously described in its monomeric format, showing a strong rotavirus neutralization activity [31]
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call