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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, izvirni znanstveni članek Opis: 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.
Najdeno v: osebi
Ključne besede: nanobodies, synthetic phage display library, in vitro panning
Objavljeno: 17.08.2016; Ogledov: 2569; Prenosov: 176
 Polno besedilo (4,32 MB) |
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, objavljeni povzetek znanstvenega prispevka na konferenci Opis: A promising new approach for the detection of biomarkers in patient sera
Najdeno v: osebi
Ključne besede: AFM, diagnostic antibodies, HER2
Objavljeno: 17.08.2016; Ogledov: 2271; Prenosov: 0
Polno besedilo (854,47 KB)
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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, izvirni znanstveni članek Opis: 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
Najdeno v: osebi
Ključne besede: nanobodies, molecular dynamics, modeling, antibody solubility
Objavljeno: 11.10.2016; Ogledov: 2414; Prenosov: 181
Polno besedilo (1,95 MB) |
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10. Nanobodies against surface biomarkers enable the analysis of tumor genetic heterogeneity in uveal melanoma Patient Derived XenograftsRonan Crepin, David Gentien, Angeline Duche, Audrey Rapinat, Cecile Reyes, Fariba Nemati, Gerald Massonnet, Didier Deacaudin, Selma Djander, Sandrine Moutel, Klervi Even Desrumeaux, Nathalie Cassoux, Sophie Piperno-Neumann, Sebastian Amigorena, Franck Perez, Sergio Roman-Roman, Ario de Marco, 2017, izvirni znanstveni članek Opis: Monoclonal antibodies specific for biomarkers expressed on the surface of uveal melanoma (UM) cells would simplify the immune-capture and genomic characterization of heterogeneous tumor cells originated from patient derived xenografts (PDXs). Antibodies against four independent tumor antigens were isolated by panning a nanobody synthetic library. Such antibodies enabled flow-cytometry-based sorting of distinct cell sub-populations from UM PDXs and to analyze their genomic features. The complexity and specificity of the biochemical and genomic biomarker combinations mirrored the UM tumor polyclonality. The data showed that MUC18 is highly and universally displayed at the surface of UM cells with different genetic background and consequently represents a reliable pan-biomarker for their identification and purification. In contrast, the other three biomarkers were detected in very variable combinations in UM PDX cells. The availability of the identified nanobodies will be instrumental in developing clone-specific drug evaluation and rational clinical strategies based on accurate genomic profiling
Najdeno v: osebi
Ključne besede: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Objavljeno: 19.04.2017; Ogledov: 3203; Prenosov: 0
Polno besedilo (1,11 MB) |
