Monoclonal Antibodies Against G Protein of Spring Viremia of Carp Virus

Abstract

Part of the G protein (3094-4170 bp) of spring viremia of carp virus (SVCV) was expressed in Escherichia coli and purified by dialysis in our study. Two clones of monoclonal antibodies (MAbs 1H11 and 4B8) against G protein were generated by fusion of mouse myeloma cell line SP2/0 and spleen lymphocytes from part of G protein (3094-4170 bp) immunized mice. The results of ELISA (enzyme-linked immunosorbent assay), IFA (indirect immunofluorescent assay), and Western blot assay further demonstrated the characterizations of the two MAbs. Both 1H11 and 4B8 were specific to SVCV G protein. Ten pairs of synthesized overlapping peptides were used to identify the epitope of the MAbs. The MAbs are useful in the development of SVCV diagnostic methods.

Analytical validation of two RT-qPCR tests and detection of spring viremia of carp virus (SVCV) in persistently infected koi Cyprinus carpio

“…The latter are produced in response to SVCV infection in carp (Sulimanovic 1973, Hill et al 1975, Fijan et al 1977 and to vesiculoviruses, the closest relatives to spriviviruses (Björklund et al 1996, Dietzgen et al 2017, which show evidence of immune selection in the G protein sequence (Kelley et al 1972, Vandepol et al 1986, Munis et al 2018. The region encoded by the Q1G target sequence (pre-cleavage amino acids 184−214) does not overlap with regions encoding neutralizing epitopes predicted for SVCV (Luo et al 2014, Zhu et al 2019 or vesicular stomatis virus (VSV) (Vandepol et al 1986, Keil & Wagner 1989, Munis et al 2018. Nevertheless, viral adaptation due to immune selection or other selection pressures acting in this region may pose a potential risk for future false negative results if nucleotide changes occur in the Q1G target sequence.…”

Cyprinid viral diseases and vaccine development

“…Moreover, the indirect immunofluorescence assay and ELISA appear to cross-react with other rhabdoviruses [80], leading to possible false-positive diagnoses. mAbs are also useful tools for detecting SVCV and studying the function of virus-specific proteins [82,83]. Recently, a single-chain fragment variable antibody against SVCV has been developed using phage display technology and employed for rapid detection of SVCV [84].…”

Zebrafish as a model for Rhabdovirus infection and molecular mechanism of the immune response

“…采 用反向遗传系统验证缺失NV基因的重组SHRV病毒研 究, 结果发现缺失NV基因突变体或含有SHRV基因组 重组病毒感染斑马鱼有相同死亡率 [144] . 也有报道认为 NV基因在斑马鱼免疫应答机制中没有明显作用, 并且 SHRV病毒G蛋白可以由IHNV病毒G蛋白代替发挥功 能 [145,146] . 因此, 鱼类弹状病毒结构蛋白诱导斑马鱼免 疫应答的分子机制可能有所不同, 目前主要发现有弹 状病毒G, M, N蛋白等参与调控过程 [137,146] .…”

“…也有报道认为 NV基因在斑马鱼免疫应答机制中没有明显作用, 并且 SHRV病毒G蛋白可以由IHNV病毒G蛋白代替发挥功 能 [145,146] . 因此, 鱼类弹状病毒结构蛋白诱导斑马鱼免 疫应答的分子机制可能有所不同, 目前主要发现有弹 状病毒G, M, N蛋白等参与调控过程 [137,146] . 毒增殖 [147,148] .…”