An RNA-to-RNA pipeline for rapid antiviral antibody development

Area of Science:

• Biotechnology
• Immunology
• Virology

Background:

• Antibody therapeutic development faces challenges with recombinant protein production.
• Existing methods for antigen generation and antibody manufacturing can be time-consuming and resource-intensive.

Purpose of the Study:

• To establish a self-amplifying replicon RNA (repRNA) platform for an end-to-end RNA-to-antibody-to-RNA workflow.
• To demonstrate the platform's efficacy against enterovirus D68 (EV-D68), a pathogen lacking vaccines or treatments.

Main Methods:

• Alpacas were immunized with repRNA encoding virus-like particles to generate antibodies.
• Phage display libraries were constructed from peripheral blood mononuclear cells to identify broadly neutralizing heavy-chain-only antibodies (VHHs).
• Lead VHHs were re-encoded into repRNA for in vivo therapeutic delivery.

Main Results:

• The platform successfully identified VHH antibodies against EV-D68.
• repRNA-encoded VHHs demonstrated protective effects against EV-D68 challenge in mice.
• Cryo-electron microscopy elucidated the VHHs' capsid-binding footprint and neutralization mechanism.

Conclusions:

• The developed repRNA platform offers a modular approach for rapid discovery and delivery of antiviral biologics.
• This technology holds promise for addressing emerging infectious diseases like EV-D68.
• The study showcases a novel strategy for therapeutic antibody development bypassing traditional recombinant protein limitations.