Interference of p53:Twist1 interaction through competing nanobodies

D'Agostino, Serena; Mazzega, Elisa; Praček, Katja; Piccinin, Sara; Pivetta, Flavia; Armellin, Michela; Fortuna, Sara; Maestro, Roberta; Marco, Ario de

doi:10.1016/j.ijbiomac.2021.11.160

Cited by 5 publications

(5 citation statements)

References 47 publications

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“…Our work confirms that in vitro selection of nanobodies performed imposing stringent parameters represents a valuable modality to isolate binders specific for particular domains, such as antigen epitopes and Ab paratopes [ 34 , 35 ]. We obtained a large and structurally highly heterogeneous array of clones with specificity for a specific idiotype.…”

Section: Discussionsupporting

confidence: 74%

Nanobodies Selectively Binding to the Idiotype of a Dengue Virus Neutralizing Antibody Do Not Necessarily Mimic the Viral Epitope

et al. 2023

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Vaccination against dengue virus is challenged by the fact that a generic immune response can induce antibody-dependent-enhancement (ADE) in secondary infections. Only some antibodies targeting a quaternary epitope formed by the dimerization of the virus protein E possess sufficient neutralizing capacity. Therefore, the immunization with anti-idiotypic antibodies of neutralizing antibodies might represent a safe vaccination strategy. Starting from a large pre-immune library, we succeeded in isolating a wide set of anti-idiotypic nanobodies characterized by selective and strong binding to the paratope of the neutralizing antibody 1C10. However, the mice immunized with such constructs did not produce effective antibodies, despite at least some of them eliciting an immune response selective for the nanobody variable regions. The results suggest that complex conformational epitopes might be difficult to be recreated by anti-idiotypic structures. The selection process of the anti-idiotypic candidates might be optimized by applying epitope mapping and modeling approaches aimed at identifying the key residues that is necessary to bind to trigger selective immune response.

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Section: Discussionsupporting

confidence: 74%

Nanobodies Selectively Binding to the Idiotype of a Dengue Virus Neutralizing Antibody Do Not Necessarily Mimic the Viral Epitope

et al. 2023

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“…The same principle could be applied in the future for the selection of nanobodies specific for EV sub-populations characterized by the presence of exclusive biomarkers. In this perspective, we already demonstrated the possibility to isolate nanobodies from a phage display pre-immune library panning directly on EVs or on specific epitopes of soluble antigens [ 8 , 37 ].…”

Section: Resultsmentioning

confidence: 99%

Macroporous Epoxy-Based Monoliths Functionalized with Anti-CD63 Nanobodies for Effective Isolation of Extracellular Vesicles in Urine

Neumair

D’Ercole

March

et al. 2023

IJMS

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Extracellular vesicles (EVs) have enormous potential for the implementation of liquid biopsy and as effective drug delivery means, but the fulfilment of these expectations requires overcoming at least two bottlenecks relative to their purification, namely the finalization of reliable and affordable protocols for: (i) EV sub-population selective isolation and (ii) the scalability of their production/isolation from complex biological fluids. In this work, we demonstrated that these objectives can be achieved by a conceptually new affinity chromatography platform composed of a macroporous epoxy monolith matrix functionalized with anti-CD63 nanobodies with afflux of samples and buffers regulated through a pump. Such a system successfully captured and released integral EVs from urine samples and showed negligible unspecific binding for circulating proteins. Additionally, size discrimination of eluted EVs was achieved by different elution approaches (competitive versus pH-dependent). The physical characteristics of monolith material and the inexpensive production of recombinant nanobodies make scaling-up the capture unit feasible and affordable. Additionally, the availability of nanobodies for further specific EV biomarkers will allow for the preparation of monolithic affinity filters selective for different EV subclasses.

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“…D’agostino et al [ 128 ] selected, with a llama immune library, the D11 VHH antibody, specific to the Twist1 factor that inhibits the p53 protein. The activity of D11 VHH was tested in fibrosarcoma cells via intracellular expression of the antibody using lentiviral vectors [ 128 ]. Pastushok et al [ 129 ] developed, using an immunized Phage Display library, an anti-RAD51 scFv-Fc.…”

Section: Phage Display To Select Antibodies Against Cancermentioning

confidence: 99%

Progress on Phage Display Technology: Tailoring Antibodies for Cancer Immunotherapy

França,

Studart,

Bezerra

et al. 2023

Viruses

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The search for innovative anti-cancer drugs remains a challenge. Over the past three decades, antibodies have emerged as an essential asset in successful cancer therapy. The major obstacle in developing anti-cancer antibodies is the need for non-immunogenic antibodies against human antigens. This unique requirement highlights a disadvantage to using traditional hybridoma technology and thus demands alternative approaches, such as humanizing murine monoclonal antibodies. To overcome these hurdles, human monoclonal antibodies can be obtained directly from Phage Display libraries, a groundbreaking tool for antibody selection. These libraries consist of genetically engineered viruses, or phages, which can exhibit antibody fragments, such as scFv or Fab on their capsid. This innovation allows the in vitro selection of novel molecules directed towards cancer antigens. As foreseen when Phage Display was first described, nowadays, several Phage Display-derived antibodies have entered clinical settings or are undergoing clinical evaluation. This comprehensive review unveils the remarkable progress in this field and the possibilities of using clever strategies for phage selection and tailoring the refinement of antibodies aimed at increasingly specific targets. Moreover, the use of selected antibodies in cutting-edge formats is discussed, such as CAR (chimeric antigen receptor) in CAR T-cell therapy or ADC (antibody drug conjugate), amplifying the spectrum of potential therapeutic avenues.

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Interference of p53:Twist1 interaction through competing nanobodies

Cited by 5 publications

References 47 publications

Nanobodies Selectively Binding to the Idiotype of a Dengue Virus Neutralizing Antibody Do Not Necessarily Mimic the Viral Epitope

Nanobodies Selectively Binding to the Idiotype of a Dengue Virus Neutralizing Antibody Do Not Necessarily Mimic the Viral Epitope

Macroporous Epoxy-Based Monoliths Functionalized with Anti-CD63 Nanobodies for Effective Isolation of Extracellular Vesicles in Urine

Progress on Phage Display Technology: Tailoring Antibodies for Cancer Immunotherapy

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