Summary auto-generated
Researchers produced three mouse monoclonal antibodies (MAbs) against potato virus X (PVX), a filamentous plant virus with significant agricultural impact. Using various immunological techniques, the team characterized these antibodies (21XB4, 21XD2, and 23XA5) and demonstrated they recognized at least two distinct epitopes on the viral coat protein. Importantly, two of the MAbs (21XD2 and 23XA5) bound to a 68-amino-acid N-terminal fragment of the PVX coat protein, indicating this region is exposed at the virus surface and highly immunogenic. All three MAbs showed high specificity for PVX and did not cross-react with other potato viruses. In double antibody sandwich ELISA assays, the MAbs successfully detected PVX at concentrations as low as 10-20 ng/ml and effectively identified the virus in potato leaf and tuber extracts. The MAbs also recognized multiple PVX isolates tested, including mild mosaic and ringspot strains, suggesting they target epitopes common across different PVX variants. These findings established practical tools for reliable serological detection of PVX in plant samples.
Key findings
- Three mouse monoclonal antibodies to PVX recognized at least two distinct epitopes, with MAbs 21XD2 and 23XA5 binding to a linear epitope in the N-terminal 68-amino-acid region of the coat protein
- The N-terminus of PVX coat protein is exposed at the virus surface and is highly immunogenic, as confirmed by both monoclonal and polyclonal antibody reactivity
- Monoclonal antibodies detected PVX in purified preparations and plant samples at sensitivities of 10-20 ng/ml in DAS ELISA, successfully identifying virus in potato leaves and tubers at high dilutions
- All three MAbs recognized multiple PVX isolates including Russian, Finnish mild mosaic, and ringspot strains, indicating they bind common epitopes across PVX variants
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Abstract
1 Laboratory of Immunodiagnostics, Järva-Jaani Collective Farm, 202823 Paide Rajoon, Estonian S.S.R.
2 Department of Molecular Genetics, Institute of Chemical Physics and Biophysics, Academy of Sciences of the Estonian S.S.R., 200026 Tallinn, Akadeemia 23, Estonian S.S.R.
3 Estonian Biocenter, 202400 Tartu, Kingissepa 14/16, Estonian S.S.R.
and4 Institute of Organic Chemistry, Department of Bioorganic Chemistry, Academy of Sciences of the Ukrainian S.S.R., Kiev 94, Ukrainian S.S.R., U.S.S.R.
A panel of mouse monoclonal antibodies (MAbs) against potato virus X (PVX) was obtained and three of these which had high affinity to the antigen were characterized in detail. These three antibodies defined two epitopes on PVX and recognized native virus, viral coat protein and denatured viral coat protein in various immunological assays. Two of the MAbs and rabbit anti-PVX polyclonal antibodies bound to the 68 amino acid N-terminal peptide of the PVX coat protein. This implies that the N terminus of the PVX coat protein is exposed at the virus surface and forms a highly immunogenic antigenic determinant. In double antibody sandwich (DAS) ELISA, MAbs and their horseradish peroxidase conjugates reacted with PVX at 10 to 20 ng/ml. Monoclonal antibodies to PVX reacted with virus in potato leaves and tubers and detected the virus in DAS ELISA in various combinations, including in combination with polyclonal antibodies.