HTRF Resource Library

Development of an HTRF® Antibody Screening Assay for Cell Line Development

By: Esohe Idusogie

OncoMed Pharmaceuticals

Abstract

In the production of monoclonal antibodies there is increasing demand to generate high producing cell lines and this usually requires screening large number of transfectants.  Selecting the appropriate screening assay is important as the throughput of the assay is critical and a potential bottle neck in the process.  Traditional antibody screening methods such as enzyme linked immunosorbent assays (ELISAs) are time consuming even in an automated format.  The homogeneous time resolved fluorescence (HTRF) technology by CISBIO presents an opportunity for developing cost effective high throughput antibody screening assays.  Using this technology, we developed a screening assay using XL665 labeled protein A as the acceptor and Europium Cryptate labeled anti-mouse antibody as the donor fluorophore. This assay is simple, homogeneous, fast and completed in a few steps.  In this study we demonstrate the assay specifically detects IgG in CHO cell culture fluid and is consistent in ranking the top antibody producing clones.  Also, a high number of transfectants (~6000/ day) can been screened with ease in a manually operated environment.  This assay has better throughput and is more cost effective when compared with other available antibody screening methods.