Nanobodies are of high interest in many fields of medicine and biotechnology, although nanobody discovery has been limited by laborious screening techniques. Here, we demonstrate the successful adaptation of linking B cell receptor to antigen specificity through sequencing (LIBRA-seq) to immunized alpacas for the rapid identification of antigen-specific nanobodies, derived from heavy-chain antibodies. We validated LIBRA-seq for nanobody discovery (nbLIBRA-seq) in two different disease settings. First, we identified over 300 antigen-specific heavy-chain antibodies against human transferrin receptor (TfR1) from a single alpaca blood sample. Second, we tested the efficiency of nbLIBRA-seq with multiple antigens in the screening library. Using fusion glycoproteins from the related respiratory syncytial virus (RSV) and human metapneumovirus (hMPV), 1,125 antigen-specific heavy-chain-expressing B cells were recovered. Our results illustrate the potential of nbLIBRA-seq to rapidly identify antigen-specific heavy-chain antibodies for a range of diverse targets, a capability that will be critical for the efficient development of nanobody-based therapeutics.
Sabina E. W. Leonard, Perry T. Wasdin, Katherine Webb et al.· Cell Reports Methods· 0 citations
Seasonal influenza A viruses cause significant global morbidity each year. Although vaccination remains the primary preventive strategy, effectiveness is often reduced by antigenic drift. This challenge is particularly pronounced for influenza A(H3N2), which has required eight vaccine updates over the past decade. Here, we present a computational framework to engineer broadly reactive influenza A(H3N2) vaccines, using protein language models to generate novel hemagglutinin (HA) sequences and a machine learning model to predict antigenic distance from circulating strains. In a proof-of-concept study, seven HA candidates designed using sequence data from 2013–2018 were evaluated in mice against contemporary and subsequently circulating viruses. Two candidates elicited protective levels of reactive antibodies, robust H3-specific antibody-secreting cell responses, and cross-neutralization against contemporary clades and drifted 2019-2020 strains. These findings demonstrate that an integrated generation–selection strategy can enhance vaccine coverage across current and future A(H3N2) seasons and may be applicable to other influenza subtypes.
Victoria R. Howard, James D. Allen, Matthew H. Thomas et al.· bioRxiv· 0 citations
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