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4. NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodiesJean-Christophe Rain, Laetitia Ligat, Ario de Marco, Emilie Lemesre, Laura Keller, Virginie Bernard, Nicolas Bery, Sandrine Moutel, Gilles Fevre, Aurelien Olichon, Franck Perez, 2016, original scientific article Abstract: In vitro selection of antibodies allows to obtain highly functional binders, rapidly and at
lower cost. Here, we describe the first fully synthetic phage display library of humanized llama
single domain antibody (NaLi-H1: Nanobody Library Humanized 1). Based on a humanized synthetic
single domain antibody (hs2dAb) scaffold optimized for intracellular stability, the highly diverse
library provides high affinity binders without animal immunization. NaLi-H1 was screened following
several selection schemes against various targets (Fluorescent proteins, actin, tubulin, p53, HP1).
Conformation antibodies against active RHO GTPase were also obtained. Selected hs2dAb were
used in various immunoassays and were often found to be functional intrabodies, enabling tracking
or inhibition of endogenous targets. Functionalization of intrabodies allowed specific protein
knockdown in living cells. Finally, direct selection against the surface of tumor cells produced
hs2dAb directed against tumor-specific antigens further highlighting the potential use of this
library for therapeutic applications. Found in: osebi Keywords: nanobodies, synthetic phage display library, in vitro panning Published: 17.08.2016; Views: 4259; Downloads: 238
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5. A Nano-Imunoassay based on fluorescence and atomic force spectroscopy for the detection of circulating cancer biomarkers: The case of HER2 positive breast cancerArio de Marco, Alessandro Bosco, Pietro Parisse, Elena Ambrosetti, Loredana Casalis, Elsa Tagliabue, 2016, published scientific conference contribution abstract Abstract: A promising new approach for the detection of biomarkers in patient sera Found in: osebi Keywords: AFM, diagnostic antibodies, HER2 Published: 17.08.2016; Views: 3769; Downloads: 0
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6. Molecular dynamics simulations and docking enable to explore the biophysical factors controlling the yields of engineered nanobodiesArio de Marco, Miguel Soler, Sara Fortuna, 2016, original scientific article Abstract: Nanobodies (VHHs) have proved to be valuable substitutes of conventional antibodies for molecular
recognition. Their small size represents a precious advantage for rational mutagenesis based on
modelling. Here we address the problem of predicting how Camelidae nanobody sequences can tolerate
mutations by developing a simulation protocol based on all-atom molecular dynamics and wholemolecule
docking. The method was tested on two sets of nanobodies characterized experimentally
for their biophysical features. One set contained point mutations introduced to humanize a wild type
sequence, in the second the CDRs were swapped between single-domain frameworks with Camelidae
and human hallmarks. The method resulted in accurate scoring approaches to predict experimental
yields and enabled to identify the structural modifications induced by mutations. This work is a
promising tool for the in silico development of single-domain antibodies and opens the opportunity to
customize single functional domains of larger macromolecules Found in: osebi Keywords: nanobodies, molecular dynamics, modeling, antibody solubility Published: 11.10.2016; Views: 4023; Downloads: 240
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10. Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymerArio de Marco, Milica Popovic, Suzana Matijasevic, Goran Brajuskovic, Aleksandra Korac, Lidija Filipovic, Milica Spasojevic, Radivoje Prodanovic, 2022, original scientific article Found in: osebi Keywords: nanobodies, immunopurification, extracellular vesicles, polymers Published: 18.03.2022; Views: 911; Downloads: 0
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