Phenotypic lentivirus screens to identify functional single domain antibodies

Florian I. Schmidt, Leo Hanke, Benjamin Morin, Rebeccah Brewer, Vesna Brusic, Sean P.J. Whelan, Hidde L. Ploegh

Research output: Contribution to journalArticlepeer-review

40 Scopus citations


Manipulation of proteins is key in assessing their in vivo function. Although genetic ablation is straightforward, reversible and specific perturbation of protein function remains a challenge. Single domain antibody fragments, such as camelid-derived VHHs, can serve as inhibitors or activators of intracellular protein function, but functional testing of identified VHHs is laborious. To address this challenge, we have developed a lentiviral screening approach to identify VHHs that elicit a phenotype when expressed intracellularly. We identified 19 antiviral VHHs that protect human A549 cells from lethal infection with influenza A virus (IAV) or vesicular stomatitis virus (VSV), respectively. Both negative-sense RNA viruses are vulnerable to VHHs uniquely specific for their respective nucleoproteins. Antiviral VHHs prevented nuclear import of viral ribonucleoproteins or mRNA transcription, respectively, and may provide clues for novel antiviral reagents. In principle, the screening approach described here should be applicable to identify inhibitors of any pathogen or biological pathway.

Original languageEnglish
Article number16080
JournalNature microbiology
Issue number8
StatePublished - Jun 20 2016


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