Cloning Mouse B Cells for Antibody Production Using Akadeum’s Novel High-Throughput Mouse B Cell Isolation Microbubbles
November 2022 Share
In a Head-to-Head Comparison Akadeum’s Mouse B Cell Isolation Microbubbles Generated 32% More Positive Antigen-Specific Antibodies Compared to Magnetic-Based Procedures
Abbratech compared Akadeum’s novel Mouse B Cell Isolation Kit with buoyancy-activated cell sorting (BACSTM) microbubbles to a negative selection magnetics-based kit for isolation of B cells from fresh splenocytes to determine which process would be incorporated into their mouse B cell cloning workflow. After enrichment, samples were cultured under various conditions and the production of antigen-specific antibodies was assessed via ELISA.
Akadeum’s Mouse B Cell Isolation Kit has a higher throughput than the magnetics-based kit, allowing Abbratech to triple the number of samples that could be processed at a given time. Most importantly for Abbratech’s purposes, Akadeum’s microbubbles were gentler compared to the magnetics-based kit: samples isolated using Akadeum’s Microbubbles yielded an average of 32% more positive wells than samples isolated using the magnetics-based kit. Thus, giving Abbratech a greater number of cells to work with to produce antibodies for their customers in a third of the time of their previous workflow.
“From the number of cells used in this work, we were able to see that Akadeum kit was clearly making a difference in the health of cells and allow for better downstream processing.”
- Results show higher throughput, processing three times the number of samples in a third of the time
- B cells were 32% more likely to survive and produce antibodies specific to the target of interest
Figure 1: Enrichment antibody cocktails.
Comparison of magnetics-based kit and Akadeum’s mouse B cell enrichment negative selection antibody cocktails.
Figure 2: Akadeum B cell isolation results in a higher frequency of antigen-specific cells ready for downstream sub-cloning.
To determine which isolation method yielded the highest number of antigen-specific clones and to optimize culture conditions, splenocytes were isolated from mice immunized with keyhole limpet hemocyanin (KLH). Samples were then enriched in a head-to-head comparison using either the Akadeum Mouse B Cell Isolation Kit or a comparable magnetics-based kit. After enrichment samples were cultured for 6 days at two cell densities in two different media formulations. Akadeum’s kit had a higher throughput, processing three times the number of samples in a third of the times, compared to the magnetics-based kit. Under optimized culture condition, B cells isolated using Akadeum’s microbubbles were 32% more likely to produce antibodies specific to the target of interest than those isolated by magnetics.
“Akadeum’s kits allowed us to increase the number of samples that we can handle at one time.”
Overall, Akadeum’s Microbubble cell isolation technology proved to be cost-effective and time-saving thereby allowing Abbratech to improve their mouse B cell cloning pipeline.
Founded in 2020, Abbratech is a comprehensive antibody development company, focused on developing more consistent, highly precise antibodies in a fraction of the time. Abbratech offers several services including antigen and project design, recombinant antibody development, antibody optimization, and antibody production.
About Akadeum Life Sciences
Akadeum has developed the next generation in cell separation using buoyancy-activated cell sorting (BACS) microbubbles. These microbubbles have revolutionized the cell separation, allowing scientists to develop treatments in a fraction of the time of previous workflows. Experience the microbubble difference for yourself! Book a meeting with our expert scientific staff to discuss your application, or shop our Mouse B Cell Isolation and other Microbubble Products online at www.akadeum.com.