Abstract 3168: High-throughput hybridoma monoclonality characterization using the Celigo™ Image Cytometer

Abstract Hybridomas are hybrid cells produced by fusing an antibody producing B cell with a myeloma cell to produce a specific monoclonal antibody. In the process of generating stable clones, developers are often required to perform several rounds of subcloning by plating cultures with a density of a single cell per well. The wells with a single colony are then selected to produce the target antibodies. Traditionally, this lengthy process was heavily relied on researchers with extensive training and experience; however, even experienced researchers may suffer from human error of selecting clones that are not truly monoclonal, thus potentially delaying antibody development and increasing costs. In this work, we demonstrate a high-throughput solution that has successfully established monoclonality of our hybridoma by employing the Celigo™ Image Cytometer (Revvity Health Sciences, Inc., Lawrence, MA) with the robotic arm accessory to screen potential candidates for downstream antibody production under a GMP environment. The Celigo™ is a plate-based, whole well, high-throughput image cytometry system with brightfield and fluorescence imaging capabilities. In combination with the robotic arm, automation can be implemented to further streamline the monoclonality workflow, and subsequently reduce subcloning personnel time by 25-50%, while also minimizing human handling errors by at least 10-fold. Following imaging acquisition, an in-house monoclonality analytical software was utilized to provide concrete documentation for every clone selected for production. Lastly, we demonstrated that time for hybridoma development in clone confirmation screening as well as subcloning could be further improved by incorporating ClonePix technology. Citation Format: Mong-Shang Lin, Carolina Franco Nitta, Mackenzie Pierce, Bo Lin, Leo L. Chan, Jessie Ni. High-throughput hybridoma monoclonality characterization using the Celigo™ Image Cytometer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3168.