Conventional and In Silico-Based Approaches for the Optimization of Nanobody Biophysical Characteristics

A special issue of Biomolecules (ISSN 2218-273X).

Deadline for manuscript submissions: closed (15 July 2024) | Viewed by 1376

Special Issue Editor


E-Mail Website
Guest Editor
Lab of Environmental and Life Sciences, University of Nova Gorica, 5000 Rožna Dolina, Nova Gorica, Slovenia
Interests: nanobodies; protein functionalization; protein quality control; pre-immune libraries; alternative scaffolds

Special Issue Information

Dear Colleagues,

Nanobodies have become a particularly appreciated class of reagents to be used in many different fields of basic research, diagnostics, and therapy. A clear advantage of these recombinant antibodies is the simplicity with which they can be engineered into improved versions of the binders originally recovered by panning. Apart from random mutagenesis, their short sequences make them particularly suitable for rational design and the resulting variants can easily be tested and compared in terms of their biophysical characteristics. This opportunity has been exploited, in combination with improvements relative to their expression (e.g., purification and combination in different formats and fusions), to produce high amounts of nanobodies and nanobody-fusion proteins with desiderable features and functions suitable for final applications. Nanobodies are indeed now a sort of model for testing new approaches, both related to conventional chemical and biochemical methodologies and driven by in silico algorithms, that aim at the more rapid and safer optimization of pre-existing binders. This collection wishes to gather innovative ideas conceived for the achievement of new paradigms in the field of nanobody technology (rational design, innovative functionalization strategies, selection procedures, expression and purification optimization, methods to improve selectivity, epitope recognition, or specific antigen conformations).

Prof. Dr. Ario de Marco
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biomolecules is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • nanobodies
  • modeling
  • screening strategies
  • random mutagenesis
  • rational engineering
  • expression conditions

Benefits of Publishing in a Special Issue

  • Ease of navigation: Grouping papers by topic helps scholars navigate broad scope journals more efficiently.
  • Greater discoverability: Special Issues support the reach and impact of scientific research. Articles in Special Issues are more discoverable and cited more frequently.
  • Expansion of research network: Special Issues facilitate connections among authors, fostering scientific collaborations.
  • External promotion: Articles in Special Issues are often promoted through the journal's social media, increasing their visibility.
  • e-Book format: Special Issues with more than 10 articles can be published as dedicated e-books, ensuring wide and rapid dissemination.

Further information on MDPI's Special Issue polices can be found here.

Published Papers (1 paper)

Order results
Result details
Select all
Export citation of selected articles as:

Research

10 pages, 1246 KiB  
Article
A Practical Guide for the Quality Evaluation of Fluobodies/Chromobodies
by Urša Štrancar, Claudia D’Ercole, Lucia Cikatricisová, Mirna Nakić, Matteo De March and Ario de Marco
Biomolecules 2024, 14(5), 587; https://doi.org/10.3390/biom14050587 - 15 May 2024
Viewed by 1077
Abstract
Background: Fluorescent proteins (FPs) are pivotal reagents for flow cytometry analysis or fluorescent microscopy. A new generation of immunoreagents (fluobodies/chromobodies) has been developed by fusing recombinant nanobodies to FPs. Methods: We analyzed the quality of such biomolecules by a combination of gel filtration [...] Read more.
Background: Fluorescent proteins (FPs) are pivotal reagents for flow cytometry analysis or fluorescent microscopy. A new generation of immunoreagents (fluobodies/chromobodies) has been developed by fusing recombinant nanobodies to FPs. Methods: We analyzed the quality of such biomolecules by a combination of gel filtration and SDS-PAGE to identify artefacts due to aggregation or material degradation. Results: In the SDS-PAGE run, unexpected bands corresponding to separate fluobodies were evidenced and characterized as either degradation products or artefacts that systematically resulted in the presence of specific FPs and some experimental conditions. The elimination of N-terminal methionine from FPs did not impair the appearance of FP fragments, whereas the stability and migration characteristics of some FP constructs were strongly affected by heating in loading buffer, which is a step samples undergo before electrophoretic separation. Conclusions: In this work, we provide explanations for some odd results observed during the quality control of fluobodies and summarize practical suggestions for the choice of the most convenient FPs to fuse to antibody fragments. Full article
Show Figures

Figure 1

Back to TopTop