Monoclonal antibodies against the structural proteins of viral haemorrhagic septicaemia virus isolates

Abstract

Five VHSV isolates from different host species and Spanish geographical locations and three viral haemorrhagic septieaemia virus (VHSV) international reference serotypes (Fj, FT and 23-75) were studied by several characterized monoclonal antibodies (MAbs) including a neutralizing MAb to four structural proteins of VHSV. We report here the lack of reaction between anti-Ml and some of the isolates of VHSV and the homogeneity of most of the isolates with respect to the MAbs tested. The reagents obtained will improve diagnostic tests which currently use polyclonal antibodies.

Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

“…15-mer peptides from the gpG of VHSV in solid phase were incubated with purified IgG isolated from murine ascites containing anti-gpG MAbs. Means and standard dev.iations from 2 different experiments are shown MAbs were I10 and C10 (Mourton, et al 1990), IPlH3 (Lorenzen et al 1990), lHlO (Sanz & Coll 1992, Sanz et al 1993) and 3FlA2 (Lorenzen unp'ubl.). MAbs 1H10, C10 and 3FlA2 were neutralizing.…”

“…The final boost was carrled out by intravenous injection of purified soluble gpGs protein (10 pg per injection). Spleen cells from immunized mice were fused with the myeloma cell lines SP2 or P3-X63-Ag.653 myeloma line (Sanz et al 1993). Screening of the hybridoma supernatants was performed by indirect ELISA on gpG4 coated plates and by flow cytometry (FACS) on VHSV-infected EPC cells.…”

“…Screening of the hybridoma supernatants was performed by indirect ELISA on gpG4 coated plates and by flow cytometry (FACS) on VHSV-infected EPC cells. Culture supernatants from the selected clones were concentrated by ammonium sulfate precipitation or purified by protein A-Sepharose (Pharmacia) following methods described before (Sanz et al 1993). The concentrated samples were applied onto precoated nitrocellulose membrane strips (ImmunoType Kit, Sigma) to determine the MAbs isotypes by following the procedure recommended by the manufacturers of the kit.…”

Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus

“…Similar results were also reported in studies developing MAbs agaist a grouper iridovirus (Shi et al, 2003), haemocytes of scallop (Xing and Zhan, 2005), lymphocystis disease virus (Cheng et al, 2006), S. maximus rhabdovirus (Zhou et al, 2006) in aquaculture research, respectively. Concerning fish rhabdoviruses, previous reports on MAbs focused mainly on virus detection using the antigen capture ELISA, sandwich ELISA and immunoperoxidase techniques (Schultz et al, 1985;Mourton et al, 1992;Sanz et al, 1992;Reno and Lannan, 1992;Reschova et al, 2007). With the development of modern immunological techniques, many new diagnosis and characterization methods of virus pathogens have been established in virtue of MAbs.…”

Recombinant protein fragments from haemorrhagic septicaemia rhabdovirus stimulate trout leukocyte anamnestic responses in vitro

“…The gels were stained with Coomassie blue and/or silver nitrate (Bio-Rad silver staining kit) or subjected to immunoblotting with anti-VHSV PAbs or MAbs (Sanz et al, 1993). Protein content was measured by A~a ~ determination, using e values of 1 for protein extracts, of 1.6 for G protein and of 0.4 for N protein, calculated by the PHYSCHEM program of the PC gene package (Intelligenetics).…”