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1.
Selection criteria and engineering of nanobodies for oncological applications
Ario De Marco, invited lecture at foreign university

Abstract: Lecture relative to the biotechnological features of nanobodies
Found in: ključnih besedah
Summary of found: ...nanobodies, phage display, antibody engineering, panning...
Keywords: nanobodies, phage display, antibody engineering, panning
Published: 14.07.2016; Views: 1276; Downloads: 0
.pdf Fulltext (22,12 KB)

2.
NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Jean-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
Summary of found: ...Here, we describe the first fully synthetic phage display library of humanized llama single domain antibody...
Keywords: nanobodies, synthetic phage display library, in vitro panning
Published: 17.08.2016; Views: 1645; Downloads: 133
.pdf Fulltext (4,32 MB)

3.
Nanobodies for research & clinical applications
Ario De Marco, invited lecture at foreign university

Abstract: Nanobody technology evolved rapidly in the last years and now is available for applications in basic sciences as well as in diagnostics and therapy
Found in: ključnih besedah
Summary of found: ...nanobodies, antibody functionalization, phage display panning, recombinant antibody production...
Keywords: nanobodies, antibody functionalization, phage display panning, recombinant antibody production
Published: 11.10.2016; Views: 1518; Downloads: 0
.pdf Fulltext (773,47 KB)

4.
Advantages offered by the nanobody format when highly-specific reagents are needed for biotechnological applications
Ario De Marco, invited lecture at foreign university

Found in: ključnih besedah
Summary of found: ...Antibody panning, pre-immune phage display library, nanobody, tumor marker, bacterial display...
Keywords: Antibody panning, pre-immune phage display library, nanobody, tumor marker, bacterial display
Published: 18.05.2017; Views: 1031; Downloads: 0
.pdf Fulltext (2,23 MB)

5.
A comprehensive nanobody platform to answer to diversified biotechnological needs
Ario De Marco, invited lecture at foreign university

Found in: ključnih besedah
Summary of found: ...nanobodies, phage display, fluorescent recombinant antibodies...
Keywords: nanobodies, phage display, fluorescent recombinant antibodies
Published: 11.10.2018; Views: 470; Downloads: 0
.pdf Fulltext (234,23 KB)

6.
Recombinant nanobodies as cheap and customizable reagents for unicellular algae detection
Elisa Mazzega, Ario De Marco, Marina Cabrini, Alfred Beran, published scientific conference contribution abstract

Abstract: At the present, the identification of planktonic species in coastal water mainly relies on light microscopy observations. This kind of analyses is performed by highly trained personnel, requires lab equipment and long processing time. High-throughput and easy-to-perform methods are instead highly needed for routine costal and ballast water monitoring. Immuno-reagents are widely employed in the medical field for routine diagnostics, where they provide the necessary sensitivity and specificity, as for example for cancer subtype characterization. Reagents of similar grade are so far not widely available for both diagnostics and basic research of microalgae. We describe the first successful isolation of a single-domain antibody (nanobody or VHH) from a pre-immune library, its engineering into application-ready reagents, and its inexpensive production as recombinant fusion protein. Alexandrium minutum was chosen as a model organism to test the feasibility of the procedure. The procedure foresees the panning of a pre-immune phage library of VHHs that was used for in vitro selection against directly the target cells. Monoclonal nanobodies specific for A. minutum cells were identified and optimized for recombinant production as fusion with fluorescent proteins in bacterial hosts. Such fluorescently-tagged VHHs were validated by immunofluorescence and cytofluorimetry for their selectivity by testing unicellular algal species that can be found in the same environment of A. minutum. Two nanobodies were found to be highly specific for the target cells, were able to bind also cysts of A. minutum and they gave no cross-reaction, even for a not-toxic strain of the closely related A. tamutum. Different tags can be then fused to the selected nanobodies and used instead of the fluorescent proteins to obtain a reagent immediately applicable to further techniques, such as cell Enzyme Linked Immuno Sorbent Assay (ELISA) or biosensor surface functionalization. The newly produced reagents can be applied for direct whole-cell detection in seawater, bypassing the need of cell processing required for DNA or RNA diagnostics, and can be used for both alive and fixed cells, guaranteeing the possibility to check old samples and to perform confirmatory morphological studies.
Found in: ključnih besedah
Summary of found: ...procedure foresees the panning of a pre-immune phage library of VHHs that was used for...
Keywords: Nanobodies, toxic algae, detection, recombinant reagent, naive library, phage display
Published: 21.12.2018; Views: 571; Downloads: 0
.pdf Fulltext (4,97 MB)

7.
8.
Phage display and Shiga toxin neutralizers
Robert Alvin Bernedo Navarro, Tomomasa Yano, 2016, review article

Found in: ključnih besedah
Keywords: Shiga toxins, phage display
Published: 27.05.2019; Views: 245; Downloads: 0
.pdf Fulltext (642,20 KB)

9.
Heptapeptides as potential neutralizers for Shiga toxin 1 cytotoxicity.
Robert Alvin Bernedo-Navarro, Mayara Mayele Miyachiro, Yano Tomomasa, unpublished conference contribution

Found in: ključnih besedah
Summary of found: ...Shiga toxins, peptide phage display, neutralizers...
Keywords: Shiga toxins, peptide phage display, neutralizers
Published: 05.06.2019; Views: 376; Downloads: 0
.pdf Fulltext (1,22 MB)

10.
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