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https://www.readbyqxmd.com/read/29766584/direct-immunoassay-for-facile-and-sensitive-detection-of-small-molecule-aflatoxin-b1-based-on-nanobody
#1
Yanfei Shen, Deng Pan, Guanghui Li, Huizhen Hu, Huaijia Xue, Mingming Zhang, Min Zhu, Xue Gong, Yuanjian Zhang, Yakun Wan
Aflatoxin B₁ (AFB₁), one of the most toxic mycotoxins, is classified as a group I carcinogen and ubiquitous in various foods and agriproducts. Thus, accurate and sensitive determination of AFB₁ is of great significance to meet the criteria of food safety. Direct detection of AFB₁ is difficult by monoclonal antibody (mAb) with large molecular size (~150 kD) since the target is too small to produce a detectable signal change. Herein, by combining the electrochemical properties of nanomaterials and the advantages of nanobodies, we developed a direct, highly selective and sensitive electrochemical immunosensor for small molecule detection...
May 15, 2018: Chemistry: a European Journal
https://www.readbyqxmd.com/read/29763554/template-catalyzed-disulfide-conjugation-of-monoclonal-antibodies-using-a-natural-amino-acid-tag
#2
Jeremy D King, Yuelong Ma, Yi-Chui Kuo, Krzysztof P Bzymek, Leah H Goodstein, Kassondra Meyer, Roger E Moore, Desiree Crow, David Colcher, Gagandeep Singh, David A Horne, John C Williams
The high specificity and favorable pharmacological properties of monoclonal antibodies (mAbs) have prompted significant interest in re-engineering this class of molecules to add novel functionalities for enhanced therapeutic and diagnostic potential. Here, we used the high affinity, meditope-Fab interaction to template and drive the rapid, efficient, and stable site-specific formation of a disulfide bond. We demonstrate that this template-catalyzed strategy provides a consistent and reproducible means to conjugate fluorescent dyes, cytotoxins, or "click" chemistry handles to meditope-enabled mAbs (memAbs) and memFabs...
May 15, 2018: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/29748336/xpo7-is-a-broad-spectrum-exportin-and-a-nuclear-import-receptor
#3
Metin Aksu, Tino Pleiner, Samir Karaca, Christin Kappert, Heinz-Jürgen Dehne, Katharina Seibel, Henning Urlaub, Markus T Bohnsack, Dirk Görlich
Exportins bind cargo molecules in a RanGTP-dependent manner inside nuclei and transport them through nuclear pores to the cytoplasm. CRM1/Xpo1 is the best-characterized exportin because specific inhibitors such as leptomycin B allow straightforward cargo validations in vivo. The analysis of other exportins lagged far behind, foremost because no such inhibitors had been available for them. In this study, we explored the cargo spectrum of exportin 7/Xpo7 in depth and identified not only ∼200 potential export cargoes but also, surprisingly, ∼30 nuclear import substrates...
May 10, 2018: Journal of Cell Biology
https://www.readbyqxmd.com/read/29733750/delivery-of-alx-0171-by-inhalation-greatly-reduces-respiratory-syncytial-virus-disease-in-newborn-lambs
#4
Alejandro Larios Mora, Laurent Detalle, Jack M Gallup, Albert Van Geelen, Thomas Stohr, Linde Duprez, Mark R Ackermann
Respiratory syncytial virus (RSV) is a common cause of acute lower respiratory disease in infants and young children worldwide. Currently, treatment is supportive and no vaccines are available. The use of newborn lambs to model hRSV infection in human infants may provide a valuable tool to assess safety and efficacy of new antiviral drugs and vaccines. ALX-0171 is a trivalent Nanobody targeting the hRSV fusion (F) protein and its therapeutic potential was evaluated in newborn lambs infected with a human strain of RSV followed by daily ALX-0171 nebulization for 3 or 5 consecutive days...
May 7, 2018: MAbs
https://www.readbyqxmd.com/read/29726815/cryo-em-structure-of-the-adenosine-a-2a-receptor-coupled-to-an-engineered-heterotrimeric-g-protein
#5
Javier García-Nafría, Yang Lee, Xiaochen Bai, Byron Carpenter, Christopher G Tate
The adenosine A2A receptor (A2A R) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein GS . Here we determine the structure by electron cryo-microscopy (cryo-EM) of A2A R at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-GS , the βγ subunits and nanobody Nb35. Most regions of the complex have a resolution of ~3.8 Å or better. Comparison with the 3.4 Å resolution crystal structure shows that the receptor and mini-GS are virtually identical and that the density of the side chains and ligand are of comparable quality...
May 4, 2018: ELife
https://www.readbyqxmd.com/read/29710823/nanobody-technology-for-mycotoxin-detection-in-the-field-of-food-safety-current-status-and-prospects
#6
REVIEW
Ting He, Jiang Zhu, Yao Nie, Rui Hu, Ting Wang, Peiwu Li, Qi Zhang, Yunhuang Yang
Mycotoxins, which are toxic, carcinogenic, and/or teratogenic, have posed a threat to food safety and public health. Sensitive and effective determination technologies for mycotoxin surveillance are required. Immunoassays have been regarded as useful supplements to chromatographic techniques. However, conventional antibodies involved in immunoassays are difficult to be expressed recombinantly and are susceptible to harsh environments. Nanobodies (or VHH antibodies) are antigen-binding sites of the heavy-chain antibodies produced from Camelidae...
April 29, 2018: Toxins
https://www.readbyqxmd.com/read/29707108/glioblastoma-specific-anti-tufm-nanobody-for-in-vitro-immunoimaging-and-cancer-stem-cell-targeting
#7
Neja Samec, Ivana Jovcevska, Jure Stojan, Alja Zottel, Mirjana Liovic, Michael P Myers, Serge Muyldermans, Jernej Šribar, Igor Križaj, Radovan Komel
Glioblastoma multiforme (GBM) is the most common and lethal form of brain tumor. The prognosis for patients remains poor, despite the combination of new preoperative and intraoperative neuroimaging, radical surgery, and recent advances in radiotherapy and chemotherapy. To improve GBM therapy and patient outcome, sustained drug delivery to glioma cells is needed, while minimizing toxicity to adjacent neurons and glia cells. This might be achieved through an anti-proteomic approach based on nanobodies, the single-domain antigen-binding fragments of heavy-chain antibodies of the camelid adaptive immune system...
April 3, 2018: Oncotarget
https://www.readbyqxmd.com/read/29706656/the-constitutive-activity-of-the-virally-encoded-chemokine-receptor-us28-accelerates-glioblastoma-growth
#8
Raimond Heukers, Tian Shu Fan, Raymond H de Wit, Jeffrey R van Senten, Timo W M De Groof, Maarten P Bebelman, Tonny Lagerweij, Joao Vieira, Sabrina M de Munnik, Laura Smits-de Vries, Jody van Offenbeek, Afsar Rahbar, Diane van Hoorick, Cecilia Söderberg-Naucler, Thomas Würdinger, Rob Leurs, Marco Siderius, Henry F Vischer, Martine J Smit
Glioblastoma (GBM) is the most aggressive and an incurable type of brain cancer. Human cytomegalovirus (HCMV) DNA and encoded proteins, including the chemokine receptor US28, have been detected in GBM tumors. US28 displays constitutive activity and is able to bind several human chemokines, leading to the activation of various proliferative and inflammatory signaling pathways. Here we show that HCMV, through the expression of US28, significantly enhanced the growth of 3D spheroids of U251- and neurospheres of primary glioblastoma cells...
April 30, 2018: Oncogene
https://www.readbyqxmd.com/read/29674997/development-by-genetic-immunization-of-monovalent-antibodies-against-human-vasoactive-intestinal-peptide-receptor-1-vpac1-new-innovative-and-versatile-tools-to-study-vpac1-receptor-function
#9
Xavier Peyrassol, Toon Laeremans, Vannessa Lahura, Maja Debulpaep, Hassan El Hassan, Jan Steyaert, Marc Parmentier, Ingrid Langer
Multi-membrane spanning proteins, such as G protein-coupled receptors (GPCRs) and ion channels, are extremely difficult to purify as native proteins. Consequently, the generation of antibodies that recognize the native conformation can be challenging. By combining genetic immunization, phage display, and biopanning, we identified a panel of monovalent antibodies (nanobodies) targeting the vasoactive intestinal peptide receptor 1 (VPAC1) receptor. The nine unique nanobodies that were classified into four different families based on their CDR3 amino acid sequence and length, were highly specific for the human receptor and bind VPAC1 with moderate affinity...
2018: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/29670611/advances-in-biomarker-guided-therapy-for-pediatric-and-adult-onset-neuroinflammatory-disorders-targeting-chemokines-cytokines
#10
REVIEW
Michael R Pranzatelli
The concept and recognized components of "neuroinflammation" are expanding at the intersection of neurobiology and immunobiology. Chemokines (CKs), no longer merely necessary for immune cell trafficking and positioning, have multiple physiologic, developmental, and modulatory functionalities in the central nervous system (CNS) through neuron-glia interactions and other mechanisms affecting neurotransmission. They issue the "help me" cry of neurons and astrocytes in response to CNS injury, engaging invading lymphoid cells (T cells and B cells) and myeloid cells (dendritic cells, monocytes, and neutrophils) (adaptive immunity), as well as microglia and macrophages (innate immunity), in a cascade of events, some beneficial (reparative), others destructive (excitotoxic)...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29658806/nanobodies-and-cancer-current-status-and-new-perspectives
#11
Alessandro Allegra, Vanessa Innao, Demetrio Gerace, Doriana Vaddinelli, Andrea Gaetano Allegra, Caterina Musolino
Reducing the dimension of antigen-binding proteins to an only immunoglobulin domain has been one of the objectives of antibody manufacturing. Heavy chain antibodies were encountered while attempting to separate the blood serum proteins of dromedaries. Later the term "nanobodies" (Nbs) was introduced. The advantageous features of Nbs comprise little immunogenicity, stability at low/high pH, capacity to target antigens that are less antigenic, and, lastly, easy capability to be used for therapy against tumor cells...
April 16, 2018: Cancer Investigation
https://www.readbyqxmd.com/read/29621251/ultrastructural-localisation-of-protein-interactions-using-conditionally-stable-nanobodies
#12
Nicholas Ariotti, James Rae, Nichole Giles, Nick Martel, Emma Sierecki, Yann Gambin, Thomas E Hall, Robert G Parton
We describe the development and application of a suite of modular tools for high-resolution detection of proteins and intracellular protein complexes by electron microscopy (EM). Conditionally stable GFP- and mCherry-binding nanobodies (termed csGBP and csChBP, respectively) are characterized using a cell-free expression and analysis system and subsequently fused to an ascorbate peroxidase (APEX) enzyme. Expression of these cassettes alongside fluorescently labelled proteins results in recruitment and stabilisation of APEX, whereas unbound APEX nanobodies are efficiently degraded by the proteasome...
April 5, 2018: PLoS Biology
https://www.readbyqxmd.com/read/29603451/exploring-the-role-of-post-translational-modifications-in-regulating-%C3%AE-synuclein-interactions-by-studying-the-effects-of-phosphorylation-on-nanobody-binding
#13
Farah El Turk, Erwin De Genst, Tim Guilliams, Bruno Fauvet, Mirva Hejjaoui, Justin Di Trani, Anass Chiki, Anthony Mittermaier, Michele Vendruscolo, Hilal A Lashuel, Christopher M Dobson
Intracellular deposits of α-synuclein in the form of Lewy bodies are major hallmarks of Parkinson's disease (PD) and a range of related neurodegenerative disorders. Post-translational modifications (PTMs) of α-synuclein are increasingly thought to be major modulators of its structure, function, degradation and toxicity. Among these PTMs, phosphorylation near the C-terminus at S129 has emerged as a dominant pathogenic modification as it is consistently observed to occur within the brain and cerebrospinal fluid (CSF) of post-mortem PD patients, and its level appears to correlate with disease progression...
March 30, 2018: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/29602806/oversized-galactosides-as-a-probe-for-conformational-dynamics-in-lacy
#14
Irina Smirnova, Vladimir Kasho, Xiaoxu Jiang, Hong-Ming Chen, Stephen G Withers, H Ronald Kaback
Binding kinetics of α-galactopyranoside homologs with fluorescent aglycones of different sizes and shapes were determined with the lactose permease (LacY) of Escherichia coli by FRET from Trp151 in the binding site of LacY to the fluorophores. Fast binding was observed with LacY stabilized in an outward-open conformation ( k on = 4-20 μM-1 ·s-1 ), indicating unobstructed access to the binding site even for ligands that are much larger than lactose. Dissociation rate constants ( k off ) increase with the size of the aglycone so that K d values also increase but remain in the micromolar range for each homolog...
March 30, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29579624/modulating-ion-channel-function-with-antibodies-and-nanobodies
#15
REVIEW
Catelijne Stortelers, Carolina Pinto-Espinoza, Diane Van Hoorick, Friedrich Koch-Nolte
Immune cells express various voltage-gated and ligand-gated ion channels that mediate the influx and efflux of charged ions across the plasma membrane, thereby controlling the membrane potential and mediating intracellular signal transduction pathways. These channels thus present potential targets for experimental modulation of immune responses and for therapeutic interventions in immune disease. Small molecule drugs and natural toxins acting on ion channels have illustrated the potential therapeutic benefit of targeting ion channels on immune cells...
March 23, 2018: Current Opinion in Immunology
https://www.readbyqxmd.com/read/29577475/selectable-high-yield-recombinant-protein-production-in-human-cells-using-a-gfp-yfp-nanobody-affinity-support
#16
Matthew J Schellenberg, Robert M Petrovich, Christine C Malone, R Scott Williams
Recombinant protein expression systems that produce high yields of pure proteins and multi-protein complexes are essential to meet the needs of biologists, biochemists, and structural biologists using X-ray crystallography and cryo-electron microscopy. An ideal expression system for recombinant human proteins is cultured human cells where the correct translation and chaperone machinery are present. However, compared to bacterial expression systems, human cell cultures present several technical challenges to their use as an expression system...
March 25, 2018: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/29569425/comparative-analysis-of-nanobody-sequence-and-structure-data
#17
Laura S Mitchell, Lucy J Colwell
Nanobodies are a class of antigen-binding protein derived from camelids that achieve comparable binding affinities and specificities to classical antibodies, despite comprising only a single 15 kDa variable domain. Their reduced size makes them an exciting target molecule with which we can explore the molecular code that underpins binding specificity - how is such high specificity achieved? Here we use a novel dataset of 90 non-redundant, protein-binding nanobodies with antigen-bound crystal structures to address this question...
March 22, 2018: Proteins
https://www.readbyqxmd.com/read/29567535/nanobodies-targeting-cortactin-proline-rich-helical-and-actin-binding-regions-downregulate-invadopodium-formation-and-matrix-degradation-in-scc-61-cancer-cells
#18
Laurence Bertier, Tim Hebbrecht, Elien Mettepenningen, Natasja De Wit, Olivier Zwaenepoel, Adriaan Verhelle, Jan Gettemans
Cortactin is a multidomain actin binding protein that activates Arp2/3 mediated branched actin polymerization. This is essential for the formation of protrusive structures during cancer cell invasion. Invadopodia are cancer cell-specific membrane protrusions, specialized at extracellular matrix degradation and essential for invasion and tumor metastasis. Given the unequivocal role of cortactin at every stage of invadopodium formation, it is considered an invadopodium marker and potential drug target. We used cortactin nanobodies to examine the role of cortactin domain-specific function at endogenous protein level...
March 19, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/29562137/binding-specificities-of-nanobody%C3%A2-membrane-protein-complexes-obtained-from-chemical-cross-linking-and-high-mass-maldi-mass-spectrometry
#19
Martin Köhler, Christoph Neff, Camilo Perez, Cyrill Brunner, Els Pardon, Jan Steyaert, Gisbert Schneider, Kaspar P Locher, Renato Zenobi
The application of nanobodies as binding partners for structure stabilization in protein x-ray crystallography is taking an increasingly important role in structural biology. However, the addition of nanobodies to the crystallization matrices might complicate the optimization of the crystallization process, which is why analytical techniques to screen and characterize suitable nanobodies are useful. Here, we show how chemical cross-linking combined with high-mass matrix-assisted laser/desorption ionization mass spectrometry can be employed as a fast screening technique to determine binding specificities of intact nanobody•membrane protein complexes...
March 21, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29545609/single-and-two-photon-imaging-of-human-micrometastases-and-disseminated-tumour-cells-with-conjugates-of-nanobodies-and-quantum-dots
#20
Fernanda Ramos-Gomes, Julia Bode, Alyona Sukhanova, Svetlana V Bozrova, Mara Saccomano, Miso Mitkovski, Julia Eva Krueger, Anja K Wege, Walter Stuehmer, Pavel S Samokhvalov, Daniel Baty, Patrick Chames, Igor Nabiev, Frauke Alves
Early detection of malignant tumours and, especially, micrometastases and disseminated tumour cells is still a challenge. In order to implement highly sensitive diagnostic tools we demonstrate the use of nanoprobes engineered from nanobodies (single-domain antibodies, sdAbs) and fluorescent quantum dots (QDs) for single- and two-photon detection and imaging of human micrometastases and disseminated tumour cells in ex vivo biological samples of breast and pancreatic metastatic tumour mouse models expressing human epidermal growth factor receptor 2 (HER2) or carcinoembryonic antigen (CEA)...
March 15, 2018: Scientific Reports
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