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101.
Whole-cell biopanning with a synthetic phage display library of nanobodies enabled the recovery of follicle-stimulating hormone receptor inhibitors
Ronan 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.
Keywords: nanobodies, GPCR proteins, FSHR, panning, pre-immune antibody library
Published in RUNG: 19.10.2017; Views: 3939; Downloads: 0
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102.
Nuovi modelli di ricerca: oncologia comparativa per lo studio dei sarcomi
Ario De Marco, unpublished invited conference lecture

Keywords: Comparative oncology, sarcoma, canine model, immune-reagents
Published in RUNG: 10.10.2017; Views: 3657; Downloads: 0
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103.
104.
105.
Acting of folding effectors to improve recombinant protein yields and functional quality
Ario De Marco, 2017, independent scientific component part or a chapter in a monograph

Keywords: recombinant proteins, chaperones, osmolytes, protein aggregation
Published in RUNG: 24.05.2017; Views: 4277; Downloads: 0
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106.
107.
Nanobodies as biotechnological tools
Ario De Marco, invited lecture at foreign university

Keywords: nanobodies, pre-immune phage library, recombinant expression, biomarkers, tumors
Published in RUNG: 17.05.2017; Views: 3649; Downloads: 0
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108.
Biotechnological applications of nanobody pre-immune libraries
Ario De Marco, invited lecture at foreign university

Keywords: nanobodies, pre-immune libraries, in vitro selection, biomarkers, recombinant proteins
Published in RUNG: 16.05.2017; Views: 3822; Downloads: 0
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109.
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
Keywords: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Published in RUNG: 03.05.2017; Views: 4855; Downloads: 0
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110.
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
Keywords: nanobodies, uveal melanoma, patient derived xenografts, MUC18, membrane surface biomarkers, panning, tumor polyclonality
Published in RUNG: 19.04.2017; Views: 5490; Downloads: 0
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