Immunization for Generation of Hybridoma Antibodies Specifically Reacting with Plants Infected with a Mycoplasmalike Organism (MLO) and Their Use in Detection of MLO Antigens

Abstract

Employing a procedure by using mice neonatally injected with nontarget antigens present in immunogen preparation before immunization, mouse hybridomas that secreted antobodies 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 plant pathogenic MLO were selected (...)