Emergent Technologies for Improved Single B Cell Screening

Revolutionizing B Cell Screening Workflows

Recent advancements in single B cell screening technologies have transformed antibody discovery workflows. Traditional approaches include hybridoma technology and single B cell screening, which remain reliable. However, emergent technologies like LIBRA-seq and NanoPen platforms are improving the efficiency and scalability of antibody discovery processes.¹

LIBRA-seq: Linking B Cell Receptor to Antigen Specificity Through Sequencing

LIBRA-seq is a next-generation sequencing-based technology that links the sequences of B cell receptors (BCRs) to their antigen specificity using DNA barcoding. In a typical workflow, B cells are incubated with DNA-barcoded antigens and subsequently sorted via flow cytometry. The sorted cells are encapsulated in emulsion droplets for amplification and sequencing. This method enables high-throughput mapping of paired VH/VL genes to antigen specificity, offering high precision in identifying candidate monoclonal antibodies (mAbs).

This technology has been successfully applied to infectious disease research. For example, LIBRA-seq identified broadly neutralizing antibodies against HIV-1 by probing B cells from infected donors. These antibodies demonstrated cross-reactivity and high neutralizing activity against multiple HIV strains, showcasing the potential for targeting diverse viral variants. Similarly, LIBRA-seq has been used to identify SARS-CoV-2-neutralizing antibodies with high specificity for viral epitopes, including those shared between SARS-CoV and SARS-CoV-2.

Related: Hybridoma Sequencing

NanoPen Technology: Streamlining Antibody Discovery with Nanofluidics

NanoPen technology, utilized in platforms like the Berkeley Lights Beacon, isolates individual plasma cells in nanoliter-sized chambers, enabling high-throughput analysis of antibody-secreting cells. Each chamber houses secreted antibodies for subsequent evaluation of binding specificity, ligand-blocking activity, and other functional properties.

A recent study compared the performance of the Beacon platform to conventional methods for identifying SARS-CoV-2-neutralizing antibodies.² The NanoPen system isolated hundreds of antigen-specific B cells from convalescent COVID-19 patients. These cells were evaluated for ACE2-blocking activity, resulting in the discovery of multiple potent antibodies. Notably, this platform accelerated the antibody discovery process, condensing weeks of traditional workflows into days.

In addition to SARS-CoV-2, the Beacon platform has been used to discover antibodies with cross-species binding properties, such as human and cynomolgus antigens. It has also been adapted for the identification of single-domain antibodies (VHH) and anti-idiotypic antibodies for specialized applications.

Related: Single B Cell Screening

Functional Screening and Emerging Technologies

Functional screening evaluates antibodies for specific bioactivities, such as cell signaling inhibition, stability, or epitope blocking, early in the discovery process. For instance, technologies like CytoMine have been employed to identify antibodies with CD3 agonist activity before sequencing. Combining such capabilities with LIBRA-seq or NanoPen workflows could enable the discovery of antibodies with superior therapeutic potential.

Innovative platforms like FLUIDOT offer additional advantages by analyzing antibodies from plasma cells and other B cell subtypes. These technologies could further enhance the selection of antibodies with diverse epitopes, maximizing therapeutic efficacy.

Machine Learning and Computational Insights

Machine learning is poised to revolutionize antibody discovery by leveraging sequencing data to predict desirable antibody properties. LIBRA-seq-generated datasets, which map antibody sequences to their antigen specificities, could train models to identify novel antibodies with high affinity, specificity, and functional activity. Similarly, machine learning tools have already been employed to predict human-like antibody qualities, facilitating the humanization of therapeutic antibodies derived from animal models.

Combining experimental methods with computational approaches will optimize antibody discovery processes, enabling rapid responses to emerging diseases. As sequencing technologies evolve, their integration with machine learning will be instrumental in accelerating therapeutic development. By adopting advanced screening technologies and computational tools, the biotech industry can address challenges in antibody discovery with greater speed and precision, offering new opportunities for combating infectious diseases and beyond.

At Biointron, we are dedicated to accelerating antibody discovery, optimization, and production. Our team of experts can provide customized solutions that meet your specific research needs. Contact us to learn more about our services and how we can help accelerate your research and drug development projects.

Our High-throughput Fully Human Antibody Discovery Platform integrates Cyagen’s HUGO-Ab™ mice with Biointron’s AbDrop™ microdroplet-based single B cell screening. This powerful combination accelerates the discovery and development of fully human antibodies, reducing the time from target identification to therapeutic candidate to just three months. Learn more about the service here.

References:

1. Schardt, J. S., Sivaneri, N. S., & Tessier, P. M. (2024). Monoclonal Antibody Generation Using Single B Cell Screening for Treating Infectious Diseases. Biodrugs, 38(4), 477. https://doi.org/10.1007/s40259-024-00667-0 

2. Zost, S. J., Gilchuk, P., Chen, R. E., Case, J. B., Reidy, J. X., Trivette, A., Nargi, R. S., Sutton, R. E., Suryadevara, N., Chen, E. C., Binshtein, E., Shrihari, S., Ostrowski, M., Chu, H. Y., Didier, J. E., MacRenaris, K. W., Jones, T., Day, S., Myers, L., . . . Carnahan, R. H. (2020). Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein. Nature Medicine, 26(9), 1422. https://doi.org/10.1038/s41591-020-0998-x