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Production of Monoclonal Antibodies to Sugarcane Yellow Leaf Virus Using Recombinant Readthrough Protein
Author(s) -
Korimbocus J.,
Preston S.,
Danks C.,
Barker I.,
Coates D.,
Boonham N.
Publication year - 2002
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1046/j.1439-0434.2002.00791.x
Subject(s) - polyclonal antibodies , antiserum , monoclonal antibody , biology , microbiology and biotechnology , virology , immunoassay , western blot , antibody , recombinant dna , virus , gene , biochemistry , immunology
Yellow leaf syndrome (YLS) of sugarcane is associated with sugarcane yellow leaf virus (SCYLV), a member of the family Luteoviridae . A fragment of the coat protein and readthrough domain of SCYLV wasexpressed in a bacterial expression system. The resulting protein was purified and used to immunize mice for monoclonal antibody (MAb) production. Two MAbs, 3A2E3 and 2F7H5, were selected following the screening of hybridoma cells using both plate‐trapped antigen enzyme‐linked immunosorbent assay (PTA‐ELISA) and tissue blot immunoassay (TBIA). These MAbs can be incorporated into the TBIA assay currently used for the routine detection of SCYLV but could not be used in triple antibody sandwich enzyme‐linked immunosorbent assay (TAS‐ELISA). The two antibodies selected have slightly different specificities. Antibody 3A2E3 gave equivalent results to a polyclonal antiserum (raised to purified virus) in comparative testing using TBIA. The MAbs produced should provide a widely available, uniform reagent for SCYLV diagnosis with the potential to help manage YLS.