Hybridoma cell lines secreting monoclonal antibodies against equine infectious anemia virus.

This research describes the production of a monoclonal antibody (1B15) against the Equine Infectious Anemia Virus (EIAV) and its application in developing an assay for detecting the virus. The antibody was generated from mice cells hypersensitized with a viral antigen (p29) and tested for its specificity.

Generation of Monoclonal Antibody

• The researchers created a monoclonal antibody named 1B15 against the EIAV. They followed a process that involved the fusion of myeloma cells (a type of cancer cell) with spleen cells from mice. These mice had been intentionally exposed to the p29 viral antigen to induce a robust immune response.
• A procedure known as the Ouchterlony double-diffusion analysis showed that the 1B15 antibody belongs to the IgG class, a common class of antibodies involved in immune response.

Confirming Antibody Specificity

• The research team used cross-over immunoelectrophoresis and disc-gel electrophoresis to confirm that the immune reaction was specific to the p29 antigen, meaning the 1B15 antibody was successfully targeting the intended antigen.

Development of RIA Assay

• With the 1B15 monoclonal antibody, the researchers created a solid-phase ‘capture’ Radioimmunoassay (RIA), which is a type of test that measures the presence of antigen (in this case, EIAV-p29) by using radioactive labeling. In this assay, the antibody was able to ‘capture’ the p29 antigen when it was present.
• The process involved binding the 1B15 antibody to the surfaces of microtitre plates. When the EIAV-p29 antigen was present, it was captured by the bound antibodies. A rabbit anti-p29 serum was then used, followed by a radioactive tracer to detect the captured antigen.
• This assay was effectively employed to detect the presence of the EIAV virus in horse serum and infected cell culture fluids, offering a potential tool for diagnosing EIAV in horses.