
==== Front
bioRxiv
BIORXIV
bioRxiv
2692-8205
Cold Spring Harbor Laboratory

10.1101/2024.08.30.610421
preprint
2
Article
Rapid synthesis and screening of natively paired antibodies against influenza hemagglutinin stem via oPool + display
Ouyang Wenhao O. http://orcid.org/0000-0002-3618-9808

Lv Huibin http://orcid.org/0000-0003-2218-0076

Liu Wenkan http://orcid.org/0009-0002-1702-3148

Mou Zongjun http://orcid.org/0000-0002-1908-617X

Lei Ruipeng http://orcid.org/0000-0002-4652-3400

Pholcharee Tossapol http://orcid.org/0000-0002-5347-1936

Wang Yiquan http://orcid.org/0000-0002-1954-9808

Dailey Katrine E.
Gopal Akshita B. http://orcid.org/0000-0002-6429-0073

Choi Danbi
Ardagh Madison R.
Talmage Logan
Rodriguez Lucia A.
Dai Xinghong http://orcid.org/0000-0002-4102-3175

Wu Nicholas C. http://orcid.org/0000-0002-9078-6697

31 8 2024
2024.08.30.610421https://creativecommons.org/licenses/by-nd/4.0/ This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License, which allows reusers to copy and distribute the material in any medium or format in unadapted form only, and only so long as attribution is given to the creator. The license allows for commercial use.
http://biorxiv.org/lookup/doi/10.1101/2024.08.30.610421
nihpp-2024.08.30.610421.pdf
ABSTRACT

Antibody discovery is crucial for developing therapeutics and vaccines as well as understanding adaptive immunity. However, the lack of approaches to synthesize antibodies with defined sequences in a high-throughput manner represents a major bottleneck in antibody discovery. Here, we presented oPool + display, which combines oligo pool synthesis and mRNA display to construct and characterize many natively paired antibodies in parallel. As a proof-of-concept, we applied oPool + display to rapidly screen the binding activity of >300 natively paired influenza hemagglutinin (HA) antibodies against the conserved HA stem domain. Structural analysis of 16.ND.92, one of the identified HA stem antibodies, revealed a unique binding mode distinct from other known broadly neutralizing HA stem antibodies with convergent sequence features. Yet, despite such differences, 16.ND.92 remained broadly reactive and conferred in vivo protection. Overall, this study not only established an experimental platform that can be applied in both research and therapeutics to accelerate antibody discovery, but also provides molecular insights into antibody responses to the influenza HA stem, which is a major target for universal influenza vaccine development.
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pmc
