11. |
12. Nanobody technology: principles & applicationsArio 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
Keywords: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Published in RUNG: 03.05.2017; Views: 4912; Downloads: 0
This document has many files! More... |
13. 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, 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
Keywords: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Published in RUNG: 19.04.2017; Views: 5564; Downloads: 0
This document has many files! More... |
14. |
15. NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodiesSandrine Moutel, Nicolas Bery, Virginie Bernard, Laura Keller, Emilie Lemesre, Ario De Marco, Laetitia Ligat, Jean-Christophe Rain, 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.
Keywords: nanobodies, synthetic phage display library, in vitro panning
Published in RUNG: 17.08.2016; Views: 5196; Downloads: 243
 Full text (4,32 MB) |
16. |
