Production of an anti-idiotypic antibody single chain variable fragment vaccine against <italic>Edwardsiella tarda</italic>

Abstract

Edwardsiella tarda is the pathogen responsible for edwardsiellosis, a serious infectious disease of freshwater and marine fish species, and currently recognized to be the species pathogenic for human. An anti-idiotypic monoclonal antibody (mAb), 1E11, has been developed. It mimics the protective epitope of E. tarda and can prevent fish from infection of E. tarda. In this study, the correct variable heavy (V H ) and variable light (V L ) genes were obtained from 1E11 by using bioinformatics methods, and a 15 amino acid (Gly 4 Ser) 3 linker was used to hold the two V domains together for the construction of V Llinker -V H form of single chain variable fragment (scFv) gene. Then, the scFv was subcloned into the vector pET28a, expressed in the Escherichia coli BL21 cells, and identified by SDS -PAGE and western blotting. Red drum (Sciaenops ocellatus L.) weighing about 50 g was subjected to challenge with different E. tarda strains after 4 weeks followed by vaccination, the mortality rates and relative percentage survival were recorded and calculated, and the survival rate of fish in the scFv subgroups was obviously higher than that of control subgroups (P < 0.01). Enzyme-linked immunosorbent assay results show that after 4 weeks of post-vaccination, the level of specific antibody in fish sera of scFv groups was significantly higher than control groups. This study indicates that the recombinant antibody scFv was successfully developed, and it may serve as an effective vaccine candidate against E. tarda.

The production of monovalent and anti-idiotype antivenom against Mesobuthus eupeus (Scorpionida: Buthidae) venom in rabbits

“…The use of anti-idiotype antibody in the control of K88 Eschershia coli (Jaradat et al, 2010), Edwardsiella tarda (Qin et al, 2010), hepatitis B (Yildrim et al, 2004, HIV-1 (Mader and Kunert, 2012), prion diseases (Venturini et al, 2009), cancer (Cerio et al, 2007) and atherosclerosis (Brito et al, 2012) were reported before.…”

“…Anti-idiotype injection can be an alternative for hyperimmunization with original antigens. Using this method, researchers have been able to produce anti-idiotype antibodies against many toxins, cancers and infectious diseases that can mimic the original antigen specificity (Boutin et al, 1994;Liu et al, 1996;Yildrim et al, 2004;Jaradet et al, 2010;Qin et al, 2010). Idiotype hypotheses and anti-idiotype performance are for cellular and humoral defense (Girdhari et al, 2006).…”

The efficacy of NP11-4-derived immunotoxin scFv-artesunate in reducing hepatic fibrosis induced by Schistosoma japonicum in mice

“…Peptide linkers that join V H and V L chains together usually vary from 10-30 amino acids in length, and the most common linker is the (Gly 4 Ser) 3 [27]. Some studies have reported that variable regions could be linked in the V L -linker-V H orientation or V H -linker-V L orientation[28], and the latter is more commonly seen. The orientations can affect expression efficiency[29], stability, and antigen binding activity.…”

A novel strategy for rapid construction of libraries of full-length antibodies highly expressed on mammalian cell surfaces

“…Commonly used restriction enzymes, such as HindIII [16] and XhoI [17], form palindromes after digestion, allowing self-ligation to occur and reducing the ligation efficiency. Two different approaches were commonly used to prevent self-ligation.…”