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Computational design of customised nanobodies for biotechnological applications
Sara Fortuna, Ario de Marco, Miguel Soler, 2016, unpublished conference contribution

Abstract: In silico modeling to improve the biophysical characteristics of recombinant single-domain antibodies
Found in: ključnih besedah
Keywords: nanobodies, modeling, protein stability, antibody humanization, molecular dynamics
Published: 26.04.2016; Views: 2769; Downloads: 0
.pdf Fulltext (2,42 MB)

Recombinant antibody production evolves into multiple options aimed at yielding reagents suitable for application-specific needs
Ario De Marco, 2015, review article

Found in: ključnih besedah
Summary of found: ...recombinant antibody, nanobodies, antibody display, neutralising antibodies, ...
Keywords: recombinant antibody, nanobodies, antibody display, neutralising antibodies
Published: 17.05.2016; Views: 2678; Downloads: 152
URL Fulltext (0,00 KB)

Selection criteria and engineering of nanobodies for oncological applications
Ario de Marco, invited lecture at foreign university

Abstract: Lecture relative to the biotechnological features of nanobodies
Found in: ključnih besedah
Keywords: nanobodies, phage display, antibody engineering, panning
Published: 14.07.2016; Views: 2602; Downloads: 0
.pdf Fulltext (22,12 KB)

NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Jean-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: ključnih besedah
Keywords: nanobodies, synthetic phage display library, in vitro panning
Published: 17.08.2016; Views: 2992; Downloads: 188
.pdf Fulltext (4,32 MB)

Molecular dynamics simulations and docking enable to explore the biophysical factors controlling the yields of engineered nanobodies
Ario 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: ključnih besedah
Keywords: nanobodies, molecular dynamics, modeling, antibody solubility
Published: 11.10.2016; Views: 2859; Downloads: 195
.pdf Fulltext (1,95 MB)

Nanobodies for research & clinical applications
Ario de Marco, invited lecture at foreign university

Abstract: Nanobody technology evolved rapidly in the last years and now is available for applications in basic sciences as well as in diagnostics and therapy
Found in: ključnih besedah
Keywords: nanobodies, antibody functionalization, phage display panning, recombinant antibody production
Published: 11.10.2016; Views: 2987; Downloads: 0
.pdf Fulltext (773,47 KB)

Is there an approach for optimizing the specific binding of antibodies to target cells?
Ario De Marco, invited lecture at foreign university

Found in: ključnih besedah
Summary of found: ... Nanobodies, avidity, modeling, off-target accumulation...
Keywords: Nanobodies, avidity, modeling, off-target accumulation
Published: 31.01.2017; Views: 2508; Downloads: 0
.pdf Fulltext (81,48 KB)

Nanobody technology: principles & applications
Ario de Marco, unpublished invited conference lecture

Found in: ključnih besedah
Summary of found: ... nanobodies, nanotechnologies, biomarker identification...
Keywords: nanobodies, nanotechnologies, biomarker identification
Published: 29.03.2017; Views: 2697; Downloads: 0
.pdf Fulltext (114,69 KB)

Nanobodies against surface biomarkers enable the analysis of tumor genetic heterogeneity in uveal melanoma Patient Derived Xenografts
Ronan 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, original scientific article

Abstract: 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
Found in: ključnih besedah
Keywords: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Published: 19.04.2017; Views: 3683; Downloads: 0
.pdf Fulltext (1,11 MB)

Nanobody technology: principles & applications
Ario De Marco, invited lecture at foreign university

Abstract: Antibodies possess unattainable capacity to bind selectively and at high affinity their cognates. For this reason they have been largely used in applications which rely on specific molecular recognition. Biosensors, nanoparticles, and even cells can be functionalized with antibodies for improving sensitivity and target specificity. However, conventional antibodies (IgGs) are large molecules (150 kDa) that are difficult to engineer. In the last years, antibody fragments have become more and more popular as an effective alternative and specifically nanobodies raised enthusiasm because of their minimal mass (14 kDa), high stability, relative similarity to human sequences, and simplified mutagenesis. Pre-immune nanobody libraries have the further advantage of enabling blind selection for antigens that can be used to discriminate between subpopulations of cells and vesicles. The panning can be performed directly on intact cells and the resulting binders are specific for the native antigen conformation
Found in: ključnih besedah
Summary of found: ...popular as an effective alternative and specifically nanobodies raised enthusiasm because of their minimal mass...
Keywords: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Published: 03.05.2017; Views: 3132; Downloads: 0
.pdf Fulltext (37,92 KB)

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