Structural and functional characterization of an anti‐West Nile virus monoclonal antibody and its single‐chain variant produced in glycoengineered plants. Issue 8 (29th June 2014)

• Record Type: Journal Article
• Authors: Lai, Huafang; He, Junyun; Hurtado, Jonathan; Stahnke, Jake; Fuchs, Anja; Mehlhop, Erin; Gorlatov, Sergey; Loos, Andreas; Diamond, Michael S.; Chen, Qiang
• Is Part Of: Plant biotechnology journal. Volume 12:Issue 8(2014:Oct.)
• Page Start: 1098
• Page End: 1107
• Publication Date: 2014-06-29
• Subjects: Plant biotechnology -- Periodicals; Plant genetic engineering -- Periodicals; 630.272
• DOI: 10.1111/pbi.12217
• Languages: English
• ISSNs: 1467-7644

Abstract

<abstract abstract-type="main" id="pbi12217-abs-0001"> <title>Summary</title> <p>Previously, our group engineered a plant‐derived monoclonal antibody (MAb pE16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed a pE16 variant consisting of a single‐chain variable fragment (scFv) fused to the heavy chain constant domains (C<sub>H</sub>) of human IgG (pE16scFv‐C<sub>H</sub>). pE16 and pE16scFv‐C<sub>H</sub> were expressed and assembled efficiently in <italic>Nicotiana benthamiana</italic> ∆XF plants, a glycosylation mutant lacking plant‐specific N‐glycan residues. Glycan analysis revealed that ∆XF plant‐derived pE16scFv‐C<sub>H</sub> (∆XFpE16scFv‐C<sub>H</sub>) and pE16 (∆XFpE16) both displayed a mammalian glycosylation profile. ∆XFpE16 and ∆XFpE16scFv‐C<sub>H</sub> demonstrated equivalent antigen‐binding affinity and kinetics, and slightly enhanced neutralization of WNV <italic>in vitro</italic> compared with the parent mammalian cell‐produced E16 (mE16). A single dose of ∆XFpE16 or ∆XFpE16scFv‐C<sub>H</sub> protected mice against WNV‐induced mortality even 4 days after infection at equivalent rates as mE16. This study provides a detailed tandem comparison of the expression, structure and function of a therapeutic MAb and its single‐chain variant produced in glycoengineered plants. Moreover, it demonstrates the development of anti‐WNV MAb therapeutic variants that are equivalent in efficacy to pE16, simpler to produce, and likely safer to use as therapeutics due to their mammalian N‐glycosylation. This platform may lead to a more robust and cost‐effective production of antibody‐based therapeutics against WNV infection and other infectious, inflammatory or neoplastic diseases.</p> </abstract>