Monoclonal antibodies specific for potato mop‐top virus, and some properties of the coat protein

Abstract

SummaryA method was devised which gave consistent yields (1–2 mg/kg leaves) of potato mop‐top furovirus (PMTV) particles. Monoclonal antibodies (MAbs) were produced and some properties of 10 of them were studied. Four MAbs readily detected PMTV isolates from six countries in Northern Europe and Japan when the isolates were trapped with polyclonal antibody; and diagnostic tests based solely on MAbs (SCR 68 to coat plates and biotin‐ or enzyme‐labelled SCR 69 to detect trapped virus) were devised. The pattern of reactions of the MAbs in ELISA and immunoblots suggested that they react with at least five different epitopes.PMTV coat protein preparations were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE) and immunoblotting. Three bands of 23.9 kd, 21.5 kd and 20.5 kd were visible in silver‐stained gels and all three reacted with PMTV specific MAbs. The relative amounts of the three bands varied between different virus preparations, but the 21.5 kd band was usually the most abundant. The three bands were probably not produced by anomalous behaviour in SDS‐PAGE. Moreover PMTV protein was readily degraded by trypsin treatment giving a band of 20.5 kd. Therefore the results suggest that PMTV coat protein sub‐units are sensitive to degradation by plant proteases. At least two degraded forms were found when purified preparations were analysed by SDS‐PAGE, and the undegraded protein was estimated to be 23.9 kd.The PMTV MAbs did not react in immunoblots with SDS‐treated coat protein preparations of beet necrotic yellow vein furovirus or Indian peanut clump furovirus.