101. 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, izvirni znanstveni članek Opis: 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.
Ključne besede: nanobodies, GPCR proteins, FSHR, panning, pre-immune antibody library
Objavljeno v RUNG: 19.10.2017; Ogledov: 3992; Prenosov: 0
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104. Quantification of Circulating Cancer Biomarkers via Sensitive Topographic Measurements on Single Binder NanoarraysElena Ambrosetti, Pamela Paoletti, Alessandro Bosco, Pietro Parisse, Denis Scaini, Elda Tagliabue, Ario De Marco, Loredana Casalis, 2017, izvirni znanstveni članek Ključne besede: Biosensors, immuno-capture, nanobodies, HER2, cancer diagostics
Objavljeno v RUNG: 22.06.2017; Ogledov: 4027; Prenosov: 211
 Celotno besedilo (4,49 MB) |
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109. Nanobody technology: principles & applicationsArio 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: 4900; Prenosov: 0
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110. 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
Ključne besede: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Objavljeno v RUNG: 19.04.2017; Ogledov: 5547; Prenosov: 0
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