Work

Targeted fusion of antibody-secreting cells: Unlocking monoclonal antibody production with hybridoma technology

Year: 2025

Type: article

Abstract: Monoclonal antibodies (mAbs) produced by hybridoma technology have extensively proved their value for therapeutic, diagnostic, and biomedical research applications, despite the reported low fusion yields between short-lived B cells and immortal myeloma cells. To improve the efficiency of this process and accelerate the development of new mAbs, we characterized and isolated antibody-secreting cells (ASCs) from the spleen of immunized mice before cell fusion. This approach resulted in a high yield of hybridoma generation by increasing the probability of successive pairing between the most suitable cell fusion partners. Specifically, we developed an optimized workflow combining Fluorescence-Activated Cell Sorting (FACS) with antibody secretion assays, using a panel of five cell-surface markers (CD3, TACI, CD138, MHC-II, and B220) that allowed us to identify a particular ASC subset with key characteristics. Such ASCs exhibited a plasmablast phenotype with high MHC-II expression and secreted high levels of Ag-specific antibodies in immunized mice. These features were also found in hybridomas, suggesting a preferential fusion of myeloma cells with this ASCs subset. Finally, the targeted electrofusion of TACI high CD138 high sorted ASCs led to a 100% fusion yield compared to a non-targeted approach. In particular, over 60% of these generated hybridomas secreted Ag-specific mAbs. Collectively, these results pave the way for a highly efficient method to produce new mAbs by cell fusion, which could facilitate hybridoma generation and expand therapeutic applications of mAbs.

Source: mAbs

DOI: 10.1080/19420862.2025.2510336

Authors: Fanny Rousseau, Catherine Menier, Patricia Brochard, Stéphanie Simon, Karla Perez-Toralla, Anne Wijkhuisen

Institutions: Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Université Paris-Saclay, CEA Paris-Saclay - Etablissement de Saclay, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

Language: English

Topic: Monoclonal and Polyclonal Antibodies Research

Subfield: Radiology, Nuclear Medicine and Imaging

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $3,575