Conformational specificity of monoclonal antibodies used in the diagnosis of tomato mosaic virus

Abstract

Ten monoclonal antibodies (McAbs) were raised to the tobamovirus, tomato mosaic virus (ToMV). Seven of the McAbs reacted with ToMV only when tested in an ELISA format, involving antibody-coated plates, which preserved the quaternary conformation of the viral protein. Two McAbs which reacted with ToMV only in an ELISA using virus-coated plates recognized an antigenic determinant present only on the disassembled form of the viral coat protein. One McAb reacted with both the assembled and dissociated forms of ToMV protein. All McAbs reacted in an identical manner with the 15 ToMV strains and isolates tested, confirming the previously established serological homogeneity of the ToMV species. Only two of the McAbs cross-reacted with two other tobamoviruses: tobacco mosaic virus (TMV) and ribgrass mosaic virus. Seven variations of ELISA were compared for their suitability to detect ToMV and TMV in tomato plants. Double antibody sandwich ELISA could be performed with a single McAb to ToMV as enough accessible epitopes are present on the virus to bind simultaneously to the coating and biotin-labelled antibody.

Evidence that cowpea aphid-borne mosaic and blackeye cowpea mosaic viruses are two different potyviruses

“…Of the five ELISA procedures using MAbs (procedures 2, 3, 4, 5 and 6; Table 1), procedure 5 was the most appropriate in revealing serological differences between isolates. Because this test requires a MAb on each side of the antigen (one as a coating antibody and one as a detecting antibody), two levels of high specificity in the detection of the virus are introduced (Dekker et al, 1987;Huguenot et al, 1989). Based on preliminary screening, 350 combinations of MAbs were selected for comparison with the 16 isolates using ELISA procedure 5.…”

Characterization of Maize Streak Virus Isolates from Different Plant Species by Polyclonal and Monoclonal Antibodies

“…None of the MAbs showed any cross-reactivity with isolates that differed from M(N)M by an SDI value greater than 4.0. Cross-reactivity studies in the tobamovirus group have shown that MAbs are generally unable to detect an antigenic relationship between two viruses separated by an SDI value of 4.0 (Briand et al, 1982;Dekker et al, 1987). …”

Monoclonal Antibodies Specific for the Hemagglutinin-Neuraminidase Protein Define Neutralizing Epitopes Specific for Newcastle Disease Virus Genotype 2.VII from Egypt.

“…Having the advantage that already staining pattern in this test may give an indication whether MAbs are directed to outer glycoproteins, the test depends on manual inspection of the plates and is more di cult to standardize; in addition use of formalin may denature sensitive epitopes. To be able to use ELISA technique for high throughput screening, investigators have used antibody-or poly L-lysine-coated plates to capture and thereby preserve the conformation of a viral protein [53,54]. Our test system relied on the lectin properties of ConA as an anchor for the antigen, a system previously shown to preserves conformational epitopes of viral glycoproteins [40,41] By using a genotype 2.VI speci c monoclonal antibody that is highly reactive by HI [55], we could demonstrate, that the ConA-ELISA preserves NDV epitopes sensitive to receptor blocking.…”