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2. 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: ključnih besedah
Keywords: nanobodies, synthetic phage display library, in vitro panning
Published: 17.08.2016; Views: 4240; Downloads: 238
 Fulltext (4,32 MB) |
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4. 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
Found in: ključnih besedah
Summary of found: ...four independent tumor antigens were isolated by panning a nanobody synthetic library. Such antibodies enabled...
Keywords: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Published: 19.04.2017; Views: 4901; Downloads: 0
Fulltext (1,11 MB) |
5. 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
Found in: ključnih besedah
Summary of found: ...between subpopulations of cells and vesicles. The panning can be performed directly on intact cells...
Keywords: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Published: 03.05.2017; Views: 4219; Downloads: 0
Fulltext (37,92 KB) |
6. Advantages offered by the nanobody format when highly-specific reagents are needed for biotechnological applicationsArio de Marco, invited lecture at foreign university Found in: ključnih besedah
Summary of found: ...Antibody panning, pre-immune phage display library, nanobody, tumor marker,...
Keywords: Antibody panning, pre-immune phage display library, nanobody, tumor marker, bacterial display
Published: 18.05.2017; Views: 3177; Downloads: 0
Fulltext (2,23 MB) |
7. Whole-cell biopanning with a synthetic phage display library of nanobodies enabled the recovery of follicle-stimulating hormone receptor inhibitorsRonan Crepin, Gianluca Veggiani, Selma Djender, Anne Beugnet, Francois Planeix, Christophe Pichon, Sandrine Moutel, Sebastian Amigorena, Franck Pérez, Nicolae Ghinea, Ario De Marco, 2017, original scientific article Abstract: Antibodies are essential reagents that are increasingly used in diagnostics and therapy. Their specificity
and capacity to recognize their native antigen are critical characteristics for their in vivo application.
Follicle-stimulating hormone receptor is a GPCR protein regulating ovarian follicular maturation and
spermatogenesis. Recently, its potentiality as a cancer biomarker has been demonstrated but no antibody
suitable for in vivo tumor targeting and treatment has been characterized so far. In this paper we describe
the first successful attempt to recover recombinant antibodies against the FSHR and that: i) are directly
panned from a pre-immune library using whole cells expressing the target receptor at their surface; ii)
show inhibitory activity towards the FSH-induced cAMP accumulation; iii) do not share the same epitope
with the natural binder FSH; iv) can be produced inexpensively as mono- or bivalent functional molecules
in the bacterial cytoplasm. We expect that the proposed biopanning strategy will be profitable to
identify useful functional antibodies for further members of the GPCR class.
Found in: ključnih besedah
Summary of found: ...nanobodies, GPCR proteins, FSHR, panning, pre-immune antibody library...
Keywords: nanobodies, GPCR proteins, FSHR, panning, pre-immune antibody library
Published: 19.10.2017; Views: 3307; Downloads: 0
Fulltext (982,69 KB) |
8. Nanobodies: flexible structures for biotechnological applicationsArio De Marco, unpublished invited conference lecture Found in: ključnih besedah
Summary of found: ...nanobodies, in vitro panning, extracellular vesicles, oncology, toxic algae...
Keywords: nanobodies, in vitro panning, extracellular vesicles, oncology, toxic algae
Published: 16.03.2018; Views: 2965; Downloads: 0
Fulltext (436,52 KB) |
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