Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody

PLoS One. 2013 Aug 15;8(8):e68772. doi: 10.1371/journal.pone.0068772. eCollection 2013.

Abstract

Plant genetic engineering, which has led to the production of plant-derived monoclonal antibodies (mAb(P)s), provides a safe and economically effective alternative to conventional antibody expression methods. In this study, the expression levels and biological properties of the anti-rabies virus mAb(P) SO57 with or without an endoplasmic reticulum (ER)-retention peptide signal (Lys-Asp-Glu-Leu; KDEL) in transgenic tobacco plants (Nicotiana tabacum) were analyzed. The expression levels of mAb(P) SO57 with KDEL (mAb(P)K) were significantly higher than those of mAb(P) SO57 without KDEL (mAb(P)) regardless of the transcription level. The Fc domains of both purified mAb(P) and mAb(P)K and hybridoma-derived mAb (mAb(H)) had similar levels of binding activity to the FcγRI receptor (CD64). The mAb(P)K had glycan profiles of both oligomannose (OM) type (91.7%) and Golgi type (8.3%), whereas the mAb(P) had mainly Golgi type glycans (96.8%) similar to those seen with mAb(H). Confocal analysis showed that the mAb(P)K was co-localized to ER-tracker signal and cellular areas surrounding the nucleus indicating accumulation of the mAb(P) with KDEL in the ER. Both mAb(P) and mAb(P)K disappeared with similar trends to mAb(H) in BALB/c mice. In addition, mAb(P)K was as effective as mAb(H) at neutralizing the activity of the rabies virus CVS-11. These results suggest that the ER localization of the recombinant mAb(P) by KDEL reprograms OM glycosylation and enhances the production of the functional antivirus therapeutic antibody in the plant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Antibodies, Viral*
  • Gene Expression*
  • Glycosylation
  • Intracellular Space
  • Mice
  • Nicotiana / genetics
  • Nicotiana / metabolism
  • Plant Cells / metabolism
  • Plantibodies / chemistry
  • Plantibodies / genetics*
  • Plantibodies / immunology
  • Plantibodies / isolation & purification
  • Plantibodies / metabolism*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • Protein Transport

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Plantibodies

Grants and funding

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ009419022013 and PJ0090622013)”; Rural Development Administration and Korea Institute of Planning & Evaluation for Technology in Food, Agriculture, Forestry & Fisheries (IPET) (111096-3); and grants (MEST) (NRF-2010-0024259) from National Research Foundation, Republic of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.