Review
Biochemistry & Molecular Biology
Mehdi Arbabi-Ghahroudi
Summary: Since the discovery of camelid heavy-chain antibodies in 1993, there has been great interest in the research and application of nanobodies. After years of effort, the first nanobody drug was approved in 2018-2019 for the treatment of rare blood clotting disorders. Despite intellectual property restrictions, nanobodies have the potential to become mainstream biotherapeutics in the future.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Review
Oncology
Yasaman Asaadi, Fatemeh Fazlollahi Jouneghani, Sara Janani, Fatemeh Rahbarizadeh
Summary: By comparing the structure and properties of camelid antibodies (VHHs) and single-chain variable fragments (scFvs), it can be determined which one is more suitable for their respective therapeutic, diagnostic, and research applications.
BIOMARKER RESEARCH
(2021)
Review
Cell Biology
Jan Gettemans, Brian De Dobbelaer
Summary: Single-domain antibodies derived from camelids and sharks, known as nanobodies, have gained attention for their high specificity and affinity binding to targets. Their key features including single-domain nature, small size, affordable high-level expression, and flexibility in altering primary structure have made them appealing for researchers and valuable in biomedical research.
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
(2021)
Article
Biochemical Research Methods
Meiniang Wang, Likun Wei, Haitao Xiang, Bingzhao Ren, Xiaopan Liu, Lin Jiang, Naibo Yang, Jiahai Shi
Summary: This study demonstrates the construction and application of a megadiverse naive sdAb library derived from non-immunized camelid animals. The library provides a rapid and adaptable platform for antibody creation, yielding high-affinity VHH candidates with therapeutic and diagnostic implications.
ANALYTICAL BIOCHEMISTRY
(2022)
Article
Virology
Daria V. Voronina, Dmitry Shcheblyakov, Irina A. Favorskaya, Ilias B. Esmagambetov, Alina S. Dzharullaeva, Amir Tukhvatulin, Olga Zubkova, Olga Popova, Vladislav Y. Kan, Alina S. Bandelyuk, Maxim M. Shmarov, Denis Y. Logunov, Boris S. Naroditskiy, Aleksandr L. Gintsburg
Summary: This study modified two potent single-domain antibodies to enhance their antiviral activity against multiple influenza subtypes, demonstrating effective prophylaxis and therapy in mice.
Article
Biochemistry & Molecular Biology
Seisho Kinoshita, Makoto Nakakido, Chinatsu Mori, Daisuke Kuroda, Jose M. M. Caaveiro, Kouhei Tsumoto
Summary: Research on VHH engineering for biotechnological and clinical applications is actively conducted to take advantage of their favorable biophysical properties. Humanization of camelid VHHs through CDR grafting with a humanized VHH in clinical trials resulted in changes to the affinity, thermal stability, and conformational structure. Mutational and back-mutational analyses on the framework regions revealed the significant contribution of bulky residues and intramolecular interactions to the properties of VHHs. Back-mutations restored intramolecular interactions, resulting in increased thermal stability and affinity, showing the potential for designing VHHs by modifying the framework region without altering the CDRs.
Review
Biochemistry & Molecular Biology
Wenyi Wang, Jumao Yuan, Changan Jiang
Summary: Nanobodies, variable domains of heavy chain-only antibodies isolated from camelids, exhibit unique features such as small size, high stability, and cryptic epitopes accessibility, making them ideal for advanced applications in plants and animals. Current applications of nanobodies in plant science and biotechnology include expression in plants, biotechnological applications, toxin and pathogen determination, and resistance against plant pathogens. Prospects and challenges of nanobody applications in plants are also discussed in this review.
PLANT MOLECULAR BIOLOGY
(2021)
Article
Oncology
Yan Yan, Xiao Cheng, Lin Li, Rumeng Zhang, Yong Zhu, Zhengsheng Wu, Keshuo Ding
Summary: The study successfully constructed HER2-nanobody and demonstrated its ability to inhibit growth and promote apoptosis of HER2-positive breast cancer cells. HER2-nanobody exerts its tumor suppressive role by inhibiting the phosphorylation of ERK and AKT.
FRONTIERS IN ONCOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Seisho Kinoshita, Makoto Nakakido, Chinatsu Mori, Daisuke Kuroda, Jose M. M. Caaveiro, Kouhei Tsumoto
Summary: Camelid single domain antibodies, known as VHHs or Nanobodies, are versatile tools for various biotechnological and clinical applications due to their favorable biophysical properties. Research on VHH engineering has shown that modifications can impact affinity and thermal stability, highlighting the importance of interactions between the framework region and CDR3 for regulating VHH characteristics.
Review
Biochemistry & Molecular Biology
Angela Donato Maia Malaquias, Livia Erika Carlos Marques, Soraya S. Pereira, Cleberson de Freitas Fernandes, Andrea Queiroz Maranhao, Rodrigo G. Stabeli, Eridan Orlando Pereira Tramontina Florean, Maria Izabel Florindo Guedes, Carla Freire Celedonio Fernandes
Summary: VHHs are small functional immunoglobulins that exhibit high tissue penetration and low immunogenicity. They provide superior antigen recognition compared to human antibodies, along with better solubility and stability. Plant-based expression systems offer versatility, cost-effectiveness, safety, and efficiency in producing VHH-related constructs.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Immunology
Eugenia A. Panova, Denis A. Kleymenov, Dmitry V. Shcheblyakov, Evgeniia N. Bykonia, Elena P. Mazunina, Alina S. Dzharullaeva, Anastasia N. Zolotar, Artem A. Derkaev, Ilias B. Esmagambetov, Ivan I. Sorokin, Evgeny V. Usachev, Anatoly N. Noskov, Igor A. Ivanov, Timofei S. Zatsepin, Sergey E. Dmitriev, Vladimir A. Gushchin, Boris S. Naroditsky, Denis Y. Logunov, Alexander L. Gintsburg
Summary: Single-domain antibodies (sdAbs, VHHs, or nanobodies) are small-sized antibodies that show promise in treating infectious and somatic diseases. Their ability to bind hard-to-reach antigenic epitopes makes them valuable for therapy. Fusion of VHH with the immunoglobulin Fc fragment enhances their neutralizing activity and half-life.
FRONTIERS IN IMMUNOLOGY
(2023)
Article
Medicine, Research & Experimental
Sanne A. M. van Lith, Fokko J. Huizing, Gerben M. Franssen, Bianca A. W. Hoeben, Jasper Lok, Sofia Doulkeridou, Otto C. Boerman, Martin Gotthardt, Paul M. P. van Bergen en Henegouwen, Johan Bussink, Sandra Heskamp
Summary: This study compared variants of CAIX-binding VHH B9 with different affinities (low and high) for SPECT imaging of CAIX expression. The results showed that B9 variants with ABD demonstrated better tumor uptake, but some uptake was CAIX-independent, limiting their use in hypoxia assessment.
MOLECULAR PHARMACEUTICS
(2022)
Article
Multidisciplinary Sciences
Jessica Hong, Hyung Joon Kwon, Raul Cachau, Catherine Z. Chen, Kevin John Butay, Zhijian Duan, Dan Li, Hua Ren, Tianyuzhou Liang, Jianghai Zhu, Venkata P. Dandey, Negin P. Martin, Dominic Esposito, Uriel Ortega-Rodriguez, Miao Xu, Mario J. Borgnia, Hang Xie, Mitchell Ho
Summary: The study constructed large dromedary camel VHH phage libraries and successfully isolated two high-affinity nanobodies that showed broad neutralization activity against SARS-CoV-2 and its variants. These nanobodies can be used for therapeutic purposes against COVID-19 variants, and the dromedary camel VHH phage libraries serve as a unique platform for rapidly isolating potent nanobodies against future emerging viruses.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Review
Biochemistry & Molecular Biology
Martin A. Rossotti, Kasandra Belanger, Kevin A. Henry, Jamshid Tanha
Summary: Single-domain antibodies have desirable properties as biologic drugs, but their sequences may increase immunogenicity risks. While most clinical trials show minimal immunogenicity, special caution is needed for highly potent sdAbs. Further research is needed to establish humanization thresholds to maximize human sequence content without compromising binding affinity or stability.
Article
Microbiology
Greg Hussack, Martin A. Rossotti, Henk van Faassen, Tomohiko Murase, Luiz Eugenio, Joseph D. Schrag, Kenneth K. -S. Ng, Jamshid Tanha
Summary: Crystal structures of camelid heavy-chain antibody variable domains (V(H)Hs) bound to fragments of the combined repetitive oligopeptides domain of Clostridiodes difficile toxin A (TcdA) revealed a spatial arrangement between VHH A20 and VHH A26. Based on this observation, a biparatopic fusion protein A20-A26 was generated, which showed improved binding affinity and neutralization potency compared to unfused A20 and A26 V(H)Hs. A20-A26 demonstrated higher potency than various control constructs. Size-exclusion chromatography-multiangle light scattering (SEC-MALS) analysis confirmed the 1:1 stoichiometry and simultaneous engagement of both A20 and A26 epitopes by A20-A26, in contrast to the varied and heterogeneous binding modes of control constructs. These results underscore the importance of molecular geometric constraints in generating highly potent antibody-based reagents.
FRONTIERS IN MICROBIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Thomas Loret, Luis Augusto Visani de Luna, Alexander Fordham, Atta Arshad, Katharine Barr, Neus Lozano, Kostas Kostarelos, Cyrill Bussy
Summary: Graphene has attracted significant attention in the material community due to its unique physicochemical properties. Graphene oxide (GO), as an oxidized derivative, shows promising potential as a drug delivery system. This study investigates the lung tolerability of two different dimensions of GO and finds that nanometric GO is more suitable for pulmonary drug delivery, as it induces milder immune and tissue responses and is cleared more easily from the respiratory tract.
Correction
Nanoscience & Nanotechnology
Thomas Kisby, Acelya Yilmazer, Kostas Kostarelos
NATURE NANOTECHNOLOGY
(2022)
Article
Oncology
Lois Gardner, Kostas Kostarelo, Parag Mallick, Caroline Dive, Marilena Hadjidemetriou
Summary: Liquid biopsy assays offer opportunities for early detection of cancer and improved management of the disease by detecting cancer-associated molecular alterations in blood. This Perspective reviews key advances in liquid biopsy-based multi-omics approaches for biomarker discovery and introduces the 'nano-omics' paradigm, which utilizes nanotechnology tools for the analysis of blood-circulating cancerome.
NATURE REVIEWS CLINICAL ONCOLOGY
(2022)
Article
Toxicology
Luis Augusto Visani de Luna, Thomas Loret, Alexander Fordham, Atta Arshad, Matthew Drummond, Abbie Dodd, Neus Lozano, Kostas Kostarelos, Cyrill Bussy
Summary: The size and dose of graphene oxide (GO) were found to impact the DNA damage it caused in mouse lungs. Both micrometric and nanometric GO induced DNA damage after a single exposure, but only during the acute phase response. After repeated exposure, only a high dose of micrometric GO resulted in long-term DNA damage. Inflammation and oxidative stress were associated with the recovery or persistence of DNA damage.
PARTICLE AND FIBRE TOXICOLOGY
(2022)
Article
Engineering, Biomedical
Paul S. Sharp, Maria Stylianou, Luis M. Arellano, Juliana C. Neves, Alfredo M. Gravagnuolo, Abbie Dodd, Katharine Barr, Neus Lozano, Thomas Kisby, Kostas Kostarelos
Summary: Graphene-based 2D nanomaterials have unique physicochemical characteristics that can be used in various biomedical applications, such as drug delivery and treatment. This study investigates the distribution of thin graphene oxide (GO) nanosheets in GBM mouse models and designs a novel nano-chemotherapeutic approach using GO sheets as platforms for drug complexation. The results demonstrate enhanced cytotoxic effects of the GO-BTZ complex in vivo.
ADVANCED HEALTHCARE MATERIALS
(2023)
Article
Nanoscience & Nanotechnology
Chen-Xia Hu, Oliver Read, Yuyoung Shin, Yingxian Chen, Jingjing Wang, Matthew Boyes, Niting Zeng, Adyasha Panigrahi, Kostas Kostarelos, Igor Larrosa, Sandra Vranic, Cinzia Casiraghi
Summary: In this study, we used liquid cascade centrifugation to obtain fractions of nanosheets with different lateral size distributions from highly concentrated graphene dispersions. The concentration, stability, size, thickness, and cytotoxicity profile were studied as a function of the initial stabilizer concentration for each fraction. Our results reveal the critical importance of the initial stabilizer concentration in determining the concentration, stability, and biocompatibility of the dispersions, regardless of the lateral size of the flakes.
ACS APPLIED NANO MATERIALS
(2022)
Article
Chemistry, Multidisciplinary
Giuseppe Di Mauro, Roberta Amoriello, Neus Lozano, Alberto Carnasciali, Daniele Guasti, Maurizio Becucci, Giada Cellot, Kostas Kostarelos, Clara Ballerini, Laura Ballerini
Summary: In neuroinflammation, astrocytes respond to pro-inflammatory cytokines (CKs) through Ca2+ signaling and play important roles in disease progression. Graphene oxide (GO) nanoparticles offer a promising approach to target astrocytes and regulate their reactivity. This study investigates the effects of CKs on astrocytic Ca2+ signaling and demonstrates the potential of GO to reverse the dysregulated cellular changes in astrocytes. The findings suggest that GO-based therapeutics may be effective in treating immune-mediated neuroinflammatory diseases.
Article
Nanoscience & Nanotechnology
Guotao Peng, Hanna M. Sinkko, Harri Alenius, Neus Lozano, Kostas Kostarelos, Lars Brautigam, Bengt Fadeel
Summary: The presence of graphene oxide in the diet alters the composition of the gut microbiome in adult zebrafish, with differences seen between wild-type and ahr2-deficient animals. Furthermore, oral exposure to graphene oxide in combination with butyrate induces AhR-dependent expression of cyp1a and homing of lck(+) cells to the gut in germ-free zebrafish. Single-cell RNA sequencing studies reveal the presence of innate lymphoid cell (ILC)-like cells in germ-free zebrafish, and a "corona" of microbial butyrate triggered by graphene oxide leads to the induction of ILC2-like cells with regulatory properties.
NATURE NANOTECHNOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Thomas Loret, Luis Augusto Visani de Luna, Matteo Andrea Lucherelli, Alexander Fordham, Neus Lozano, Alberto Bianco, Kostas Kostarelos, Cyrill Bussy
Summary: The study investigates the biological response, distribution, and bio-persistence of four different graphene-based materials (GBMs) in mouse lungs. The size, carbon to oxygen ratio, and thickness of the GBMs do not induce a strong pulmonary immune response. Neutrophils internalize and degrade small GBMs faster than macrophages, revealing their crucial role in the elimination of these materials. Large GBMs, on the other hand, have hindered degradation and long-term persistence in macrophages, leading to more damages. These findings are important for the design of safer GBMs for biomedical applications.
Review
Immunology
T. H. Rabbitts
Summary: The application of antibodies in cells, particularly intracellular antibodies, has expanded over the years. These antibodies, including fragments, can be used to perturb protein activity within cells and elicit intracellular responses. They are not only research tools, but also have potential applications as drugs and templates for drug discovery. Their ability to discriminate between closely related proteins adds a new dimension to drug screening and development.
Article
Chemistry, Multidisciplinary
Lorenza Tortella, Irene Santini, Neus Lozano, Kostas Kostarelos, Giada Cellot, Laura Ballerini
Summary: Small graphene oxide nanosheets can reversibly downregulate excitatory synapses in the central nervous system and have potential applications as therapeutic tools for neuro-disorders. This study demonstrates that the delivery of s-GO can improve neuronal survival during oxygen-glucose deprivation and protect neuronal integrity under exogenous glutamate application.
CHEMISTRY-A EUROPEAN JOURNAL
(2023)
Article
Oncology
Cristiana Barone, Roberto Orsenigo, Anna Cazzola, Elisabetta D'Errico, Arianna Patelli, Giulia Quattrini, Barbara Vergani, Silvia Bombelli, Sofia De Marco, Cristina D'Orlando, Cristina Bianchi, Biagio Eugenio Leone, Raffaella Meneveri, Andrea Biondi, Giovanni Cazzaniga, Terence Howard Rabbitts, Silvia Brunelli, Emanuele Azzoni
Summary: Despite advancements in understanding the genetics of infant acute myeloid leukemia (AML), it remains unclear why this blood cancer differs from adult AML. Previous research has shown that KMT2A gene (MLL) rearrangements are common in infants with AML, but the cell of origin is unknown. This study found that introducing the Mll-Af9 rearrangement in HSC-independent progenitors led to transplantable myeloid leukemia in mice, suggesting the potential role of embryonic hematopoiesis in disease progression.
Review
Engineering, Biomedical
D. Cardella, D. Sanchez-Guzman, T. H. Rabbitts
Summary: Intracellular antibodies have been used for the past 20 years as powerful research tools to inhibit proteins and provide specific information about protein function. They have proven useful for target validation in oncology and to inhibit undruggable targets, such as RAS mutants and LMO2. They are versatile and can invoke cell phenotypes following target engagement inside cells, and their paratope-epitope interaction is being explored to develop small molecule surrogates. Their importance lies in their flexibility for discovery research and new generation therapeutics in various clinical indications involving aberrant protein expression.
CURRENT OPINION IN BIOMEDICAL ENGINEERING
(2023)
Article
Chemistry, Multidisciplinary
Helene Cazier, Carole Malgorn, Dominique Georgin, Nathalie Fresneau, Fabrice Beau, Kostas Kostarelos, Cyrill Bussy, Stephane Campidelli, Mathieu Pinault, Martine Mayne-L'Hermite, Frederic Taran, Christophe Junot, Francois Fenaille, Antoine Sallustrau, Benoit Colsch
Summary: Research on graphene based nanomaterials has expanded in recent years due to their unique properties and potential socio-economic impact. This study proposes a novel approach combining radioimaging and mass spectrometry imaging for detecting Graphene Oxide (GO) labeled with carbon-14 in mice. The results demonstrate the advantages of this correlative imaging technique for the pre-clinical development of graphene-based biomedical applications.
Article
Chemistry, Multidisciplinary
Elisa Pati, Audrey Franceschi Biagioni, Raffaele Casani, Neus Lozano, Kostas Kostarelos, Giada Cellot, Laura Ballerini
Summary: Graphene oxide nanosheets have the potential to alter neurobiological processes and regulate glutamatergic synapses, making them a promising candidate for future neurotherapeutic developments. In a rat model of post-traumatic stress disorder (PTSD), it was found that a single injection of graphene oxide to the lateral amygdala could induce behavior remission and reduce dendritic spine densities associated with PTSD. Mechanistically, graphene oxide was found to affect the probability of release, thus modulating synaptic plasticity in the amygdala.
