Immunization for generation of hybridoma antibodies specifically reacting with plants infected with a mycoplasmalike organism (MLO) and their use in detection of MLO antigens
1990
Hsu, H.T. | Lee, I.M. | Davis, R.E. | Wang, Y.C.
Employing a procedure by using mice neonatally injected with nontarget antigens present in immunogen preparation before immunization, mouse hybridomas that secreted antibodies to a mycoplasmalike organism (MLO) isolated from tomato plants with symptoms of big bud disease were produced. Neonatal mice were injected with extracts from normal host plant tissue on days 1 and 7. When mice were 8 wk old, they were immunized with a crude antigen preparation consisting of about 10% MLO as determined by dot-blot immunoassays. Based on indirect enzyme-linked immunosorbent assay 20 hybridomas that secreted specific antibodies to the plant pathogenic MLO were selected. Two hybridoma lines were used for development of a dot-blot immunoassay technique on nitrocellulose membranes for detection of the MLO. Grinding liquid nitrogen-frozen MLO-infected midrib tissues with a pestle in a mortar was an efficient method for preparation of antigen extracts for dot-blot immunoassays.
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