Nanobodies: Natural Single-Domain Antibodies

Muyldermans, Serge

doi:10.1146/annurev-biochem-063011-092449

Cited by 1,829 publications

(1,792 citation statements)

References 152 publications

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“…To target Num1 to eisosomes, we used a GFP-αGFP nanobody targeting system (Figure 1(a)). The αGFP nanobody is a ~ 16 kD, monomeric, single domain antibody that binds GFP with high affinity [22,23]. We engineered cells to express Pil1 as an αGFP nanobody fusion (Pil1-αGFP) from the endogenous PIL1 locus.…”

Section: Resultsmentioning

confidence: 99%

The role of mitochondria in anchoring dynein to the cell cortex extends beyond clustering the anchor protein

et al. 2018

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Organelle distribution is regulated over the course of the cell cycle to ensure that each of the cells produced at the completion of division inherits a full complement of organelles. In yeast, the protein Num1 functions in the positioning and inheritance of two essential organelles, mitochondria and the nucleus. Specifically, Num1 anchors mitochondria as well as dynein to the cell cortex, and this anchoring activity is required for proper mitochondrial distribution and dynein-mediated nuclear inheritance. The assembly of Num1 into clusters at the plasma membrane is critical for both of its anchoring functions. We have previously shown that mitochondria drive the assembly of Num1 clusters and that these mitochondria-assembled Num1 clusters serve as cortical attachment sites for dynein. Here we further examine the role for mitochondria in dynein anchoring. Using a GFP-αGFP nanobody targeting system, we synthetically clustered Num1 on eisosomes to bypass the requirement for mitochondria in Num1 cluster formation. Utilizing this system, we found that mitochondria positively impact the ability of synthetically clustered Num1 to anchor dynein and support dynein function even when mitochondria are no longer required for cluster formation. Thus, the role of mitochondria in regulating dynein function extends beyond simply concentrating Num1; mitochondria likely promote an arrangement of Num1 within a cluster that is competent for dynein anchoring. This functional dependency between mitochondrial and nuclear positioning pathways likely serves as a mechanism to order and integrate major cellular organization systems over the course of the cell cycle.

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Section: Resultsmentioning

confidence: 99%

The role of mitochondria in anchoring dynein to the cell cortex extends beyond clustering the anchor protein

et al. 2018

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“…These so called heavy‐chain antibodies (hcAb) are devoid of light chains, with the functional antigen‐binding unit reduced to a single variable domain (V H H, Figure 1 a) 7, 16. These properties allow the generation of potent, recombinant V H Hs by isolating and engineering the corresponding single domain region of Camelidae B cells after immunization.…”

Section: Recombinant Antigen‐binding Proteins: Nanobodies and Othersmentioning

confidence: 99%

Nanobodies: Chemical Functionalization Strategies and Intracellular Applications

et al. 2018

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Nanobodies can be seen as next‐generation tools for the recognition and modulation of antigens that are inaccessible to conventional antibodies. Due to their compact structure and high stability, nanobodies see frequent usage in basic research, and their chemical functionalization opens the way towards promising diagnostic and therapeutic applications. In this Review, central aspects of nanobody functionalization are presented, together with selected applications. While early conjugation strategies relied on the random modification of natural amino acids, more recent studies have focused on the site‐specific attachment of functional moieties. Such techniques include chemoenzymatic approaches, expressed protein ligation, and amber suppression in combination with bioorthogonal modification strategies. Recent applications range from sophisticated imaging and mass spectrometry to the delivery of nanobodies into living cells for the visualization and manipulation of intracellular antigens.

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“…[510] The Belgian company Ablynx coined the term "nanobodies" with respect to the nanometer size range of these naturally derived antigen-binding fragments, which are also termed as single domain antibodies. [511] Nanobodies, demonstrate essential characteristics such as high specificity and affinity, [504] high solubility, stability in aqueous solutions, [512] high thermal and chemical resistance, [513] capability to be chemically modified, ease of cloning, and ease of gene manipulation [496b] which relieves some restrictions related to conventional antibodies. Additionally, low immunogenicity of nanobodies which is both attributed to their high similarity with human VH sequences [514] as well as absence of Fc fragment, made nanobodies to pave their way into biomedical applications more easily.…”

Section: Antibodiesmentioning

confidence: 99%

Synthesis, Functionalization, and Design of Magnetic Nanoparticles for Theranostic Applications

Mosayebi

Kiyasatfar

Laurent

2017

Adv Healthcare Materials

203

171

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In order to translate nanotechnology into medical practice, magnetic nanoparticles (MNPs) have been presented as a class of non‐invasive nanomaterials for numerous biomedical applications. In particular, MNPs have opened a door for simultaneous diagnosis and brisk treatment of diseases in the form of theranostic agents. This review highlights the recent advances in preparation and utilization of MNPs from the synthesis and functionalization steps to the final design consideration in evading the body immune system for therapeutic and diagnostic applications with addressing the most recent examples of the literature in each section. This study provides a conceptual framework of a wide range of synthetic routes classified mainly as wet chemistry, state‐of‐the‐art microfluidic reactors, and biogenic routes, along with the most popular coating materials to stabilize resultant MNPs. Additionally, key aspects of prolonging the half‐life of MNPs via overcoming the sequential biological barriers are covered through unraveling the biophysical interactions at the bio–nano interface and giving a set of criteria to efficiently modulate MNPs' physicochemical properties. Furthermore, concepts of passive and active targeting for successful cell internalization, by respectively exploiting the unique properties of cancers and novel targeting ligands are described in detail. Finally, this study extensively covers the recent developments in magnetic drug targeting and hyperthermia as therapeutic applications of MNPs. In addition, multi‐modal imaging via fusion of magnetic resonance imaging, and also innovative magnetic particle imaging with other imaging techniques for early diagnosis of diseases are extensively provided.

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Nanobodies: Natural Single-Domain Antibodies

Cited by 1,829 publications

References 152 publications

The role of mitochondria in anchoring dynein to the cell cortex extends beyond clustering the anchor protein

The role of mitochondria in anchoring dynein to the cell cortex extends beyond clustering the anchor protein

Nanobodies: Chemical Functionalization Strategies and Intracellular Applications

Synthesis, Functionalization, and Design of Magnetic Nanoparticles for Theranostic Applications

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