Abstract
Pathogenic animal and human viruses present a growing and persistent threat to humans worldwide. Ebola virus (EBOV) causes zoonosis in humans. Here, two structurally different anti-Ebola 13F6 antibodies, recognizing the heavily glycosylated mucin-like domain (MLD) of the glycoprotein (GP), were expressed in transgenic Nicotiana tabacum plants and designed as inexpensive and effective diagnostic antibodies against Ebola virus disease (EVD). The first was anti-EBOV 13F6 full size antibody with heavy chain (HC) and light chain (LC) (monoclonal antibody, mAb 13F6-FULL), while the second was a large single-chain (LSC) antibody (mAb 13F6-LSC). mAb 13F6-LSC was constructed by linking the 13F6 LC variable region (VL) with the HC of mAb 13F6-FULL using a peptide linker and extended to the C-terminus using the endoplasmic reticulum (ER) retention motif KDEL. Agrobacterium-mediated plant transformation was employed to express the antibodies in N. tabacum. PCR, RT-PCR, and immunoblot analyses confirmed the gene insertion, transcription, and protein expression of these antibodies, respectively. The antibodies tagged with the KDEL motif displayed high-mannose type N-glycan structures and efficient binding to EBOV-like particles (VLPs). Thus, various forms of anti-EBOV plant-derived mAbs 13F6-FULL and LSC with efficient binding affinity to EBOV VLP can be produced in the plant system.
Highlights
Ebola virus disease (EVD), known as Ebola hemorrhagic fever, is an acute, severe, and often fatal illness
Purification of monoclonal antibody (mAb) 13F6-FULL and 13F6-large single-chain (LSC) Antibodies from the Transgenic N. tabacum Plant Transgenic plant lines expressing high levels of mAb 13F6-FULL and 13F6-LSC were identified by the immunoblot assay
Infectious diseases of foreign origin have shown an increasing trend worldwide. They originate from many factors, including the movement of biological life by international travel and the Infectious diseases of foreign origin have shown an increasing trend worldwide
Summary
Ebola virus disease (EVD), known as Ebola hemorrhagic fever, is an acute, severe, and often fatal illness. The LSC antibody required only one promoter to express in plant [32]. Focusing on the production of an EBOV diagnostic antibody in this study, the full type 13F6 monoclonal antibody (mAb 13F6-FULL) and an LSC type 13F6 monoclonal antibody (mAb 13F6-LSC) were expressed in transgenic Nicotiana tabacum and their in vitro efficacy was confirmed. Our data suggested that various forms of the anti-EBOV antibodies were well expressed in and purified from the transgenic N. tabacum plant. Transgenic plant lines expressing high levels of mAb 13F6-FULL and 13F6-LSC were identified by the immunoblot assay. Purification of mAb 13F6-FULL and 13F6-LSC Antibodies from the Transgenic N. tabacum Plant Transgenic plant lines expressing high levels of mAb 13F6-FULL and 13F6-LSC were identified by the immunoblot assay. SVPLRP sseunrsfoarcger. aSmPsRssheonwsonrginratmhisck lsinheoswwneirne othviecrklalpinpeesdwweirthe tohveerthlaipnpliende wanitdhththeen tfihtitnedlintoeaasnidmtphleen1:f1ititnetdertaoctaiosnimmpoldee1l:1cuirnvtee.raIcntieoanch pmlootd, tehl ecuarsvseo.cIiantieoanchanpdlodt,itshseocaisastoiocniatpiohnasaensdardeissseoecnia. t(iDon) Tphheaskeisnaertiecsceoenns.t(aDn)t Tvhaleukeisnaertiecccaolncustlaantetd uvsainlugeas asrime cpallecu1:l1atiendteurasicntigoan smimodpelel.1T:1hienmterAabct1io3nF6m-Co,daelc.oTmhemmerAcibal1a3nFt6i--CEB, aOcVomanmtiebrocdiayl wanatsi-iEnBjeOctVed aasnatibpoodsiytivweacsonintjreoclt.eVdLaPs wa apsofisxiteivdeocnoanntrioml.mVoLbPiliwzeads PfiExeGdseonnsoarncihmipm(oabpiplirzoexdimPaEtGelyse1n5s3o0rRcUhipon t(haepspernosxoimr cahteiply).1530 RU on the sensor chip)
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