Rapid detection of bovine type II collagen-specific T-cell hybridomas

T-cell hybridomas are powerful tools in studying the fine specificities of antigen recognition by the T-cell receptor (TCR), the structure and genetic basis of the CD3-TCR complex, and the size of the TCR alpha/beta repertoire used in response to various antigens. A technical challenge in establishing T-cell hybridomas is the early identification of antigen-specific ones. We have established a rapid and efficient ELISA method for detecting antigen-specific T-cell hybridomas. Our ELISA technique significantly reduces the time and resources required for the primary screening of antigen-specific T-cell hybrids, eliminates the need of maintaining hundreds of rapidly growing nonspecific clones, and does not require the maintenance of IL-2/IL-4 dependent cell lines such as CTLL-2 or HT-2. In addition, the ELISA technique is designed to detect both types of CD4 T-cells: Th1 and Th2, by using a mixture of anti-IL-2 and anti-IL-4 monoclonal antibodies. Therefore, we believe that our ELISA technique provides a faster, less expensive, and higher throughput screening method for the early identification of antigen-specific T-cell hybridomas than the current bioassays.