English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Neisseria  adhesin A (NadA), one of the surface adhesins of  Neisseria meningitides  (NM), interacts with several cell types including human brain microvascular endothelial cells (hBMECs) and play important role in the pathogenesis. Receptor binding pockets of NadA are localized on the globular head domain (A 33  to K 69 ) and the first coiled-coil domain (L 121  to K 158 ). Here, the phage display was used to develop a variable heavy chain domain (VHH) that can block receptor binding sites of recombinant NadA (rec-NadA). A phage library displaying VHH was panned against synthetic peptides (NadA-gd A33−K69  or NadA-cc L121−K158 ), gene encoding VHH was amplified from bound phages and re-cloned in the expression vector, and the soluble VHHs containing disulfide bonds were overexpressed in the SHuffle  E. coli . From the repertoire of 96 clones, two VHHs (VHH F3 –binding NadA-gd A33−K69  and VHH G9 –binding NadA-cc L121−K158 ) were finally selected as they abrogated the interaction between rec-NadA and the cell receptor. Preincubation of NM with VHH F3  and VHH G9  significantly reduced the adhesion of NM on hBMECs  in situ  and hindered the traversal of NM across the  in-vitro  BBB model. The work presents a phage display pipeline with a single-round of panning to select receptor blocking VHHs. It also demonstrates the production of soluble and functional VHHs, which blocked the interaction between NadA and its receptor, decreased adhesion of NM on hBMECs, and reduced translocation of NM across BBB  in-vitro . The selected NadA blocking VHHs could be promising molecules for therapeutic translation.

frontiers in molecular biosciences

Single Domain Antibodies Targeting Receptor Binding Pockets of NadA Restrain Adhesion of Neisseria meningitidis to Human Brain Microvascular Endothelial Cells