Efficient Identification of Monoclonal Antibodies Against Rift Valley Fever Virus Using High-Throughput Single...

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.