30 years of nanobodies – an ongoing success story of small binders in biological research

Frecot, Desiree I.; Froehlich, Theresa; Rothbauer, Ulrich

doi:10.1242/jcs.261395

Cited by 9 publications

(4 citation statements)

References 165 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Tissue clearing was performed as in ( 19 ); samples were immunolabeled with rabbit anti- Collagen IV (ColIV; Abcam ab6586, 1:200) sheep anti-von Willebrand factor (vWf; Genetex GTX74137, 1:100), mouse anti-smooth muscle actin (SMA; Abcam amab7817, 1:200), and rabbit anti-Iba1 (WAKO 019–19,741, 1:200); secondary antibodies (Invitrogen Donkey anti-sheep, anti-rabbit and anti-mouse conjugated with Alexa 488, 555 or 647) or nanobodies ( 20 ) (Synaptic System nanobodies anti-mouse and anti-rabbit conjugated with ATTO Fluor 565 and 647 dyes) were all used at 1:200. Incubations with nanobodies required half the time than with regular antibodies.…”

Section: Methodsmentioning

confidence: 99%

Temporal bone marrow of the rat and its connections to the inner ear

Perin,

Cossellu,

Vivado

et al. 2024

Front. Neurol.

View full text Add to dashboard Cite

Calvarial bone marrow has been found to be central in the brain immune response, being connected to the dura through channels which allow leukocyte trafficking. Temporal bone marrow is thought to play important roles in relation to the inner ear, but is still largely uncharacterized, given this bone complex anatomy. We characterized the geometry and connectivity of rat temporal bone marrow using lightsheet imaging of cleared samples and microCT. Bone marrow was identified in cleared tissue by cellular content (and in particular by the presence of megakaryocytes); since air-filled cavities are absent in rodents, marrow clusters could be recognized in microCT scans by their geometry. In cleared petrosal bone, autofluorescence allowed delineation of the otic capsule layers. Within the endochondral layer, bone marrow was observed in association to the cochlear base and vestibule, and to the cochlear apex. Cochlear apex endochondral marrow (CAEM) was a separated cluster from the remaining endochondral marrow, which was therefore defined as “vestibular endochondral marrow” (VEM). A much larger marrow island (petrosal non-endochondral marrow, PNEM) extended outside the otic capsule surrounding semicircular canal arms. PNEM was mainly connected to the dura, through bone channels similar to those of calvarial bone, and only a few channels were directed toward the canal periosteum. On the contrary, endochondral bone marrow was well connected to the labyrinth through vascular loops (directed to the spiral ligament for CAEM and to the bony labyrinth periosteum for VEM), and to dural sinuses. In addition, CAEM was also connected to the tensor tympani fossa of the middle ear and VEM to the endolymphatic sac. Endochondral marrow was made up of small lobules connected to each other and to other structures by channels lined by elongated macrophages, whereas PNEM displayed larger lobules connected by channels with a sparse macrophage population. Our data suggest that the rat inner ear is surrounded by bone marrow at the junctions with middle ear and brain, most likely with “customs” role, restricting pathogen spread; a second marrow network with different structural features is found within the endochondral bone layer of the otic capsule and may play different functional roles.

show abstract

Section: Methodsmentioning

confidence: 99%

Temporal bone marrow of the rat and its connections to the inner ear

Perin,

Cossellu,

Vivado

et al. 2024

Front. Neurol.

View full text Add to dashboard Cite

show abstract

“…The overexpression of cDNA is widely used to screen gene targets and study functional genomics. In genome-scale genetic screening, people screen gene targets by constructing cDNA libraries, overexpression or introducing foreign genes, and observing phenotypic changes [ 20 , 21 , 22 ]. Since overexpression can significantly increase the expression level of specific genes, it is beneficial to directly observe the corresponding cellular superficial changes after gene expression.…”

Section: Traditional Screening Techniquesmentioning

confidence: 99%

The Current Situation and Development Prospect of Whole-Genome Screening

Yang,

Lei,

Ren

et al. 2024

IJMS

View full text Add to dashboard Cite

High-throughput genetic screening is useful for discovering critical genes or gene sequences that trigger specific cell functions and/or phenotypes. Loss-of-function genetic screening is mainly achieved through RNA interference (RNAi), CRISPR knock-out (CRISPRko), and CRISPR interference (CRISPRi) technologies. Gain-of-function genetic screening mainly depends on the overexpression of a cDNA library and CRISPR activation (CRISPRa). Base editing can perform both gain- and loss-of-function genetic screening. This review discusses genetic screening techniques based on Cas9 nuclease, including Cas9-mediated genome knock-out and dCas9-based gene activation and interference. We compare these methods with previous genetic screening techniques based on RNAi and cDNA library overexpression and propose future prospects and applications for CRISPR screening.

show abstract

“…In addition to classical human and animal antibodies, significant attention has been given to the development of antiviral nanobodies during the pandemic. This antibody format, composed of VHH domains from camelid heavy-chain-only immunoglobulins, has several important advantages, including high stability, low immunogenicity in humans, ease of constructing oligomers, and the ability to recognize epitopes inaccessible to classical antibodies [ 8 , 9 ]. Given current insights into mechanisms of virus immune evasion [ 10 , 11 ], it is conceivable that at least some of the nanobody epitopes are less prone to mutation escape compared to those of the classical antibodies.…”

Section: Introductionmentioning

confidence: 99%

Serial Llama Immunization with Various SARS-CoV-2 RBD Variants Induces Broad Spectrum Virus-Neutralizing Nanobodies

Solodkov,

Najakshin,

Chikaev

et al. 2024

Vaccines

View full text Add to dashboard Cite

The emergence of SARS-CoV-2 mutant variants has posed a significant challenge to both the prevention and treatment of COVID-19 with anti-coronaviral neutralizing antibodies. The latest viral variants demonstrate pronounced resistance to the vast majority of human monoclonal antibodies raised against the ancestral Wuhan variant. Less is known about the susceptibility of the evolved virus to camelid nanobodies developed at the start of the pandemic. In this study, we compared nanobody repertoires raised in the same llama after immunization with Wuhan’s RBD variant and after subsequent serial immunization with a variety of RBD variants, including that of SARS-CoV-1. We show that initial immunization induced highly potent nanobodies, which efficiently protected Syrian hamsters from infection with the ancestral Wuhan virus. These nanobodies, however, mostly lacked the activity against SARS-CoV-2 omicron-pseudotyped viruses. In contrast, serial immunization with different RBD variants resulted in the generation of nanobodies demonstrating a higher degree of somatic mutagenesis and a broad range of neutralization. Four nanobodies recognizing distinct epitopes were shown to potently neutralize a spectrum of omicron variants, including those of the XBB sublineage. Our data show that nanobodies broadly neutralizing SARS-CoV-2 variants may be readily induced by a serial variant RBD immunization.

show abstract

30 years of nanobodies – an ongoing success story of small binders in biological research

Cited by 9 publications

References 165 publications

Temporal bone marrow of the rat and its connections to the inner ear

Temporal bone marrow of the rat and its connections to the inner ear

The Current Situation and Development Prospect of Whole-Genome Screening

Serial Llama Immunization with Various SARS-CoV-2 RBD Variants Induces Broad Spectrum Virus-Neutralizing Nanobodies

Contact Info

Product

Resources

About