Efficient Identification of Monoclonal Antibodies Against Rift Valley Fever Virus Using High-Throughput Single Lymphocyte Transcriptomics of Immunized Mice

Ronit Rosenfeld ¹, Ron Alcalay ¹, Yfat Yahalom-Ronen ²

⁰Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel.

Antibodies (basel, Switzerland) +

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February 21, 2025

View abstract on PubMed

Summary

Researchers developed potent neutralizing antibodies against Rift Valley fever virus (RVFV) using single-cell sequencing. These antibodies target RVFV surface glycoproteins and show promise for future human therapeutics and diagnostics.

Area Of Science

Virology
Immunology
Biotechnology

Background

Rift Valley fever virus (RVFV) is a zoonotic pathogen causing significant livestock and human health threats.
Human RVF disease can manifest as febrile illness, hemorrhagic fever, retinitis, or encephalitis.
No licensed RVF vaccines or therapeutics are currently available for human use, highlighting an urgent need for interventions.

Purpose Of The Study

To identify and develop novel neutralizing antibodies targeting RVFV surface glycoproteins (Gc and Gn).
To leverage advanced single-cell sequencing technology for efficient antibody discovery.
To explore the potential of these antibodies for prophylactic and therapeutic applications against RVFV.

Main Methods

Immunization of mice with a live attenuated RVFV strain and Gc/Gn protein boosts.
Isolation and sorting of memory B cell populations from splenocytes using flow cytometry.
High-throughput single-cell sequencing of the antibody repertoire and bioinformatic analysis for B cell receptor (BCR) pair selection.

Main Results

Twenty-three recombinant monoclonal antibodies (mAbs) against RVFV were successfully generated and expressed.
Approximately half of the developed mAbs exhibited specific binding to their target antigens with high affinity.
The study demonstrated the effectiveness of single-cell sequencing for identifying potent RVFV-specific antibodies.

Conclusions

The identified antibodies are promising candidates for developing effective RVF therapeutics and studying virus neutralization.
Single-cell sequencing is a powerful and applicable technique for antibody discovery against viral surface proteins.
This research paves the way for novel interventions against Rift Valley fever virus.

Keywords:

10x Genomics RVFV Gc glycoprotein RVFV Gn glycoprotein Rift Valley fever virus (RVFV) antibody discovery antibody repertoire profiling recombinant antibody single-cell analysis