Bristol Myers Squibb, Redwood City, CA, USA
5th HTRF Symposium, Avignon, France
Large scale generation of human antibodies from transgenic mice using Hybridoma technology, requires development and application of homogenous high throughput screening assays that can be applied early in the process to help selection of antibodies with desired specificity and function. Traditionally, hybridoma supernatants from 96 well fusion plates are screened by ELISA. This is time-consuming, uses significant amounts of antigen and the requirement for washing steps slows throughput. Replacing ELISA with HTRF®, a sensitive homogenous assay that is amenable to automation, allowed us to significantly increase speed and throughput, conserve operator time, and reduce stress injuries. Since all components of the assays are in solution, HTRF® also allowed us to capture antibodies binding to a wider range of epitopes. To screen hybridomas to cell surface antigens, we typically use an FMAT-based assay. We explored the Tag-lite® technology to assess specificity and sensitivity as well as increased throughput of our cell-based assays. We tested this technology using two different GPCR receptors both for antibody binding and blocking of ligand interaction. This assay was highly specific and showed good sensitivity and robustness as compared to the FMAT assay. A great advantage of Tag-lite® is that signal is detected only when the labeled receptor is engaged in the interaction. Examples of performance of these assays applied to HTP antibody screening will be provided.