Monoclonal antibodies are the workhorse molecules of the immunotherapy field. These antibodies have been engineered to bind to immune system components and redirect immune responses to kill tumor cells, dial down autoimmune responses, or enhance the effects of other treatments. Much of the work behind production of monoclonal antibodies is rooted in the production of hybridomas, which involves identifying antigen-specific plasma/plasmablast cells (ASPCs) that produce antibodies specific to an antigen of interest and fusing these cells with myeloma cells. These fused hybridoma cell lines can be grown in large-scale culture and produce a single type of (monoclonal) antibody indefinitely under the correct conditions. The ASPC selection process can be time consuming and unpredictable, but recent advances in monoclonal antibody screening that use flow cytometry are making this process faster and more efficient.
ASPC selection has been improved by screening plasma cells (PCs) stained for B cell differentiation markers using flow cytometry. This process is limited by the surface staining step, which can negatively impact plasma cell viability and downstream hybridoma fusion success.
A recent paper by Kurosawa et al.[1] describes a novel approach to ASPC selection that does not require the use of antibodies to stain plasma cells. They stained cells with a fluorescent tracker for the endoplasmic reticulum (ER), an intracellular organelle that is significantly larger in PCs because of intracellular antibody synthesis. This ER tracker can work across species and can be used in combination with staining using a fluorescently-labeled antigen to identify ASPCs. Unlike other screening protocols, this method can be used to screen a small number cells from either tissues like lymph nodes or from peripheral blood.
Consider taking your hybridoma work to the next level by using this ER-based screening for ASPCs. Many contract research organizations specialize in hybridoma production using cutting edge methods, so consider partnering with experts to create novel monoclonal antibodies that can be used as research tools or to be tested for their therapeutic potential.
Learn More About Flow Cytometry
[1] Kurosawa N, Yoshioka M, Fujimoto R, Yamagishi F, Isobe M. Rapid production of antigen-specific monoclonal antibodies from a variety of animals. BMC Biology. 2012;10:80. doi:10.1186/1741-7007-10-80.
