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11.
12.
Nanobody technology: principles & applications
Ario De Marco, predavanje na tuji univerzi

Opis: 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
Ključne besede: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Objavljeno v RUNG: 03.05.2017; Ogledov: 4856; Prenosov: 0
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13.
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, 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
Ključne besede: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Objavljeno v RUNG: 19.04.2017; Ogledov: 5491; Prenosov: 0
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14.
Nanobodies for research & clinical applications
Ario De Marco, predavanje na tuji univerzi

Opis: Nanobody technology evolved rapidly in the last years and now is available for applications in basic sciences as well as in diagnostics and therapy
Ključne besede: nanobodies, antibody functionalization, phage display panning, recombinant antibody production
Objavljeno v RUNG: 11.10.2016; Ogledov: 4878; Prenosov: 0
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15.
NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Sandrine Moutel, Nicolas Bery, Virginie Bernard, Laura Keller, Emilie Lemesre, Ario De Marco, Laetitia Ligat, Jean-Christophe Rain, 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.
Ključne besede: nanobodies, synthetic phage display library, in vitro panning
Objavljeno v RUNG: 17.08.2016; Ogledov: 5106; Prenosov: 243
.pdf Celotno besedilo (4,32 MB)

16.
Selection criteria and engineering of nanobodies for oncological applications
Ario De Marco, predavanje na tuji univerzi

Opis: Lecture relative to the biotechnological features of nanobodies
Ključne besede: nanobodies, phage display, antibody engineering, panning
Objavljeno v RUNG: 14.07.2016; Ogledov: 4249; Prenosov: 0
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