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1.
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
Najdeno v: ključnih besedah
Povzetek najdenega: ...Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering...
Ključne besede: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Objavljeno: 03.05.2017; Ogledov: 1697; Prenosov: 0
.pdf Polno besedilo (37,92 KB)

2.
Nanobodies: towards rational design of immune-reagents
Ario De Marco, 2017, objavljeni povzetek znanstvenega prispevka na konferenci (vabljeno predavanje)

Opis: Antibodies are irreplaceable reagents in both research and clinical practice. Despite their relevance, the structural complexity of conventional mono- and polyclonal antibodies (IgG) has always been a limit for their engineering towards reagents optimized for specific applications, such as in vivo diagnostics and therapy. Furthermore, their isolation is time consuming, their production expensive, and their functionalization results often in heterogeneous macromolecule populations. These drawbacks promoted the search for both innovative antibody isolation strategies and alternative scaffolds. In vitro panning of pre-immune collections of recombinant antibody fragments allows for the simple and fast recovery of binders. Since they did not undergo somatic maturation, their affinity for targets can be insufficient but on the other hand they can be rapidly mutated by standard molecular biology techniques to generate second-generation antibodies among which to identify clones with improved characteristics. Both stochastic and rational methods have been proposed for the optimization process. Random mutagenesis followed by panning at stringent conditions has been successful used to select binders with improved physical characteristics. Rational methods try to identify in silico key residues involved in the regulation of specific antibody features, such as stability or binding affinity. The accuracy of these methods usually depends on the calculation resources. In this perspective, smaller molecules can be analyzed “better” than larger because of their restricted number of residues. Nanobodies small dimensions have been long appreciated since enable better tissue penetration, shorter clearance time, higher yields. Now it becomes evident that this characteristic makes them also optimal objects for modeling.
Najdeno v: ključnih besedah
Povzetek najdenega: ...In vitro panning of pre-immune collections of recombinant antibody fragments allows for the simple and... ...has always been a limit for their engineering towards reagents optimized for specific applications, such...
Ključne besede: recombinant antibody modeling, nanobody engineering, molecular dynamics and docking
Objavljeno: 21.03.2018; Ogledov: 1156; Prenosov: 0
.pdf Polno besedilo (638,20 KB)

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