Production and characterization of monoclonal antibodies to tissue culture adapted calf rotavirus (UK-Compton strain)
Rotaviruses are a ubiquitous group of viruses responsible for causing acute gastroenteritis in the young of humans and animals. This thesis concerns the production characterization and use of monoclonal antibodies to tissue culture adapted calf rotavirus (UK-Compton strain). Monoclonal antibodies were produced, which as a result of immunoprecipitation and Transblotting experiments were determined to be directed towards VP6, the subgroup protein of calf rotavirus. Varying the immunogen and fusion protocol proved insufficient since only antibodies directed at this extremely immunogenic protein were produced. As a result of competitive binding assays, this subgroup protein was determined to consist of at least eight different epitopes. Two of these epitopes belonged to a group-specific, non-neutralizing region, whilst four epitopes belonged to a group-specific, neutralizing region. The remaining two epitopes belonged to a type-specific, neutralizing region. None of the antibodies had greater than 75% neutralization. An ELISA was developed to use four of these monoclonal antibodies to detect and subgroup rotavirus antigens using human faecal samples collected from children in Lima, Peru between the period 1983-84, during clinical trials of the RIT 4237 rotavirus vaccine. Out of a total of 61 samples, 62% were typed as belonging to subgroup 1. This was similar to a figure obtained for subgrouping human samples in the Midlands during the same period. This reflected a change in subgroup antigenicity. During 1978-82 strains belonging to subgroup 2 had been the most prevalent. The ELISA technique developed proved to be quick, cheap, efficient and reliable, thus enabling a large number of samples to be screened simultaneously.