Immune response of heterologous recombinant antigenic protein of viral hemorrhagic septicemia virus (VHSV) in mice

Abstract

Viral hemorrhagic septicemia (VHS) is an important infectious disease in fish worldwide caused by viral hemorrhagic septicemia virus (VHSV). VHSV is the causative agent of serious systemic diseases in fish, affecting a number of teleost fish species. In this study, VHSV glycoprotein (G), including its epitope, as a subunit vaccine candidate, was expressed in tobacco plant ( Nicotiana tabacum ). The recombinant gene, VHSVG, was fused to the immunoglobulin Fc fragment and extended with the endoplasmic reticulum (ER) retention signal (KDEL) to generate VHSVG-FcK. The recombinant expression vector for VHSVG-FcK was transferred into Agrobacterium tumefaciens (LBA4404), and plant transformation was conducted N. tabacum . Polymerase chain reaction (PCR) was performed to confirm gene insertion and VHSVG-FcK protein expression was confirmed by immunoblot analysis. VHSVG-FcK protein was successfully purified from tobacco plant leaves. Furthermore, ELISA analysis showed that mice serum immunized with the plant-derived VHSVG-FcK (VHSVGP-FcK) had a high absorbance against VHSVG-FcK, indicating that the plant-derived recombinant subunit vaccine protein VHSVG-FcK can induce immune response. Taken together, this recombinant vaccine protein can be expressed in plant expression systems and can be appropriately assembled to be functional in immunogenicity.

Expression of a Large Single-Chain 13F6 Antibody with Binding Activity against Ebola Virus-Like Particles in a Plant System

“…The higher purification efficiency of both antibodies might be due to the effect of the C-terminal-fused ER retention motif KDEL. Previous studies prove that mAbs were not retained in the ER in the absence of an ER retention motif [ 25 , 26 , 47 , 48 , 49 ]. They indicated that a protein fused with the KDEL motif at the C-terminal end is retained in the ER or is returned to the ER and its production levels are also improved [ 2 , 50 , 51 , 52 ].…”

“…They indicated that a protein fused with the KDEL motif at the C-terminal end is retained in the ER or is returned to the ER and its production levels are also improved [ 2 , 50 , 51 , 52 ]. The ER is a vast subcellular location for the accumulation of proteins that do not require the proteolytic process found in the secretory pathway of intracellular and extracellular spaces [ 23 , 24 , 25 , 26 , 28 , 53 , 54 ]. The figure of Immunoblot analysis to confirm the purified fraction of mAb 13F6-FULL from transgenic N. tabacum plants for Figure 4 A was included as a Figure S2 .…”

“…To make this diagnostic test cost-effective and easily accessible, low cost production of antibodies with a strong binding affinity against the EBOV antigens is crucial. One option to ensure cost-effective production of antibodies is the use of a plant expression system without pathogenic animal contaminants, high-priced manufacturing process, and expensive culture media [ 22 , 23 , 24 , 25 , 26 ]. The advantages of plant expression systems include inexpensive inputs (sunlight, water, and nutrients), the ease of large-scale production by increasing the area located in the seed, and low technology harvest compared with mammalian expression systems [ 22 , 27 , 28 , 29 ].…”

“…The advantages of plant expression systems include inexpensive inputs (sunlight, water, and nutrients), the ease of large-scale production by increasing the area located in the seed, and low technology harvest compared with mammalian expression systems [ 22 , 27 , 28 , 29 ]. Moreover, plant expression systems are suitable for post-translational modifications of proteins, including the N -glycosylation required for biological activity, similar to mammalian expression systems [ 25 , 28 , 30 , 31 ]. The structure of the antibody should be properly modified for efficient antibody production and its assembly in the plant expression system.…”

Expression and In Vitro Function of Anti-Breast Cancer Llama-Based Single Domain Antibody VHH Expressed in Tobacco Plants

“…To purify the plant-derived anti-HER2 VHH-FcK, 200 g of leaves were homogenized with extraction buffer (37.5 mM Tris/HCl pH 7.5, 50 mM NaCl, 15 mM ethylenediaminetetra acetic acid (EDTA) pH 8.0, 75 mM sodium citrate, and 0.2% sodium thiosulfate) and centrifuged at 9000× g for 30 min at 4 • C [34]. The supernatant was filtered using Miracloth (Biosciences, La Jolla, CA, USA).…”