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https://www.readbyqxmd.com/read/29137184/cell-type-specific-metabolic-flux-analysis-a-challenge-for-metabolic-phenotyping-and-a-potential-solution-in-plants
#1
Merja T Rossi, Monika Kalde, Chaiyakorn Srisakvarakul, Nicholas J Kruger, R George Ratcliffe
Stable isotope labelling experiments are used routinely in metabolic flux analysis (MFA) to determine the metabolic phenotype of cells and tissues. A complication arises in multicellular systems because single cell measurements of transcriptomes, proteomes and metabolomes in multicellular organisms suggest that the metabolic phenotype will differ between cell types. In silico analysis of simulated metabolite isotopomer datasets shows that cellular heterogeneity confounds conventional MFA because labelling data averaged over multiple cell types does not necessarily yield averaged flux values...
November 13, 2017: Metabolites
https://www.readbyqxmd.com/read/29123178/a-nanobody-based-tracer-targeting-dpp6-for-non-invasive-imaging-of-human-pancreatic-endocrine-cells
#2
Alexander Balhuizen, Sam Massa, Iris Mathijs, Jean-Valery Turatsinze, Jens De Vos, Stéphane Demine, Catarina Xavier, Olatz Villate, Isabelle Millard, Dominique Egrise, Carmen Capito, Raphaël Scharfmann, Pieter In't Veld, Piero Marchetti, Serge Muyldermans, Serge Goldman, Tony Lahoutte, Luc Bouwens, Decio L Eizirik, Nick Devoogdt
There are presently no reliable ways to quantify endocrine cell mass (ECM) in vivo, which prevents an accurate understanding of the progressive beta cell loss in diabetes or following islet transplantation. To address this unmet need, we coupled RNA sequencing of human pancreatic islets to a systems biology approach to identify new biomarkers of the endocrine pancreas. Dipeptidyl-Peptidase 6 (DPP6) was identified as a target whose mRNA expression is at least 25-fold higher in human pancreatic islets as compared to surrounding tissues and is not changed by proinflammatory cytokines...
November 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29116505/construction-of-high-quality-camel-immune-antibody-libraries
#3
Ema Romão, Vianney Poignavent, Cécile Vincke, Christophe Ritzenthaler, Serge Muyldermans, Baptiste Monsion
Single-domain antibodies libraries of heavy-chain only immunoglobulins from camelids or shark are enriched for high-affinity antigen-specific binders by a short in vivo immunization. Thus, potent binders are readily retrieved from relatively small-sized libraries of 10(7)-10(8) individual transformants, mostly after phage display and panning on a purified target. However, the remaining drawback of this strategy arises from the need to generate a dedicated library, for nearly every envisaged target. Therefore, all the procedures that shorten and facilitate the construction of an immune library of best possible quality are definitely a step forward...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29107128/targeted-cancer-therapy-through-antibody-fragments-decorated-nanomedicines
#4
REVIEW
Abbas Alibakhshi, Fatemeh Abarghooi Kahaki, Shahrzad Ahangarzadeh, Hajar Yaghoobi, Fatemeh Yarian, Roghaye Arezumand, Javad Ranjbari, Ahad Mokhtarzadeh, Miguel de la Guardia
Active targeting in cancer nanomedicine, for improved delivery of agents and diagnose, has been reviewed as a successful way for facilitating active uptake of theranostic agents by the tumor cells. The application of a targeting moiety in the targeted carrier complexes can play an important role in differentiating between tumor and healthy tissues. The pharmaceutical carriers, as main part of complexes, can be polymeric nanoparticles, micelles, liposomes, nanogels and carbon nanotubes. The antibodies are among the natural ligands with highest affinity and specificity to target pharmaceutical nanoparticle conjugates...
November 3, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/29095961/nanobodies-targeting-norovirus-capsid-reveal-functional-epitopes-and-potential-mechanisms-of-neutralization
#5
COMPARATIVE STUDY
Anna D Koromyslova, Grant S Hansman
Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs) are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important...
November 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/29095202/simultaneous-inhibition-of-tumor-necrosis-factor-receptor-1-and-matrix-metalloproteinase-8-completely-protects-against-acute-inflammation-and-sepsis
#6
Sophie Steeland, Sara Van Ryckeghem, Jolien Vandewalle, Marlies Ballegeer, Elien Van Wonterghem, Melanie Eggermont, Johan Decruyenaere, Liesbet De Bus, Claude Libert, Roosmarijn E Vandenbroucke
OBJECTIVES: Sepsis causes very high mortality and morbidity rates and remains one of the biggest medical challenges. This study investigates whether plasma levels of both matrix metalloproteinase 8 and tumor necrosis factor receptor 1 are associated with sepsis severity and also investigates the therapeutic applicability of simultaneous inhibition of the two molecules in sepsis. DESIGN: Observational human pilot study-prospective controlled animal study. SETTING: University hospital and research laboratory...
October 31, 2017: Critical Care Medicine
https://www.readbyqxmd.com/read/29090412/improved-debulking-of-peritoneal-tumor-implants-by-near-infrared-fluorescent-nanobody-image-guidance-in-an-experimental-mouse-model
#7
Pieterjan Debie, Marian Vanhoeij, Natalie Poortmans, Janik Puttemans, Kris Gillis, Nick Devoogdt, Tony Lahoutte, Sophie Hernot
PURPOSE: Debulking followed by combination chemotherapy is currently regarded as the most effective treatment for advanced ovarian cancer. Prognosis depends drastically on the degree of debulking. Accordingly, near-infrared (NIR) fluorescence imaging has been proposed to revolutionize cancer surgery by acting as a sensitive, specific, and real-time tool enabling visualization of cancer lesions. We have previously developed a NIR-labeled nanobody that allows fast, specific, and high-contrast imaging of HER2-positive tumors...
October 31, 2017: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
https://www.readbyqxmd.com/read/29061039/quantitatively-predictable-control-of-cellular-protein-levels-through-proteasomal-degradation
#8
Wenting Zhao, Lara Pferdehirt, Laura Segatori
Protein function is typically studied and engineered by modulating protein levels within the complex cellular environment. To achieve fast, targeted, and predictable control of cellular protein levels without genetic manipulation of the target, we developed a technology for post-translational depletion based on a bifunctional molecule (NanoDeg) consisting of the antigen-binding fragment from the Camelidae species heavy-chain antibody (nanobody) fused to a degron signal that mediates degradation through the proteasome...
November 7, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29057962/tnfr1-inhibition-with-a-nanobody-protects-against-eae-development-in-mice
#9
Sophie Steeland, Sara Van Ryckeghem, Griet Van Imschoot, Riet De Rycke, Wendy Toussaint, Leen Vanhoutte, Christian Vanhove, Filip De Vos, Roosmarijn E Vandenbroucke, Claude Libert
TNF has as detrimental role in multiple sclerosis (MS), however, anti-TNF medication is not working. Selective TNF/TNFR1 inhibition whilst sparing TNFR2 signaling reduces the pro-inflammatory effects of TNF but preserves the important neuroprotective signals via TNFR2. We previously reported the generation of a Nanobody-based selective inhibitor of human TNFR1, TROS that will be tested in experimental autoimmune encephalomyelitis (EAE). We specifically antagonized TNF/TNFR1 signaling using TROS in a murine model of MS, namely MOG35-55-induced EAE...
October 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29047203/engineered-cross-reacting-nanobodies-simplify-comparative-oncology-between-humans-and-dogs
#10
E Mazzega, A de Marco
Antibodies cross-reacting with homologue antigens in different species would be essential reagents for the development of comparative oncology studies. In comparison with conventional immunoglobulin Gs, recombinant nanobodies (single-domain variable regions of heavy-chain only antibodies of Camelidae origin) can be easily isolated in vitro and engineered into a variety of reagents with optimized characteristics for different research and clinical applications. We recovered an anti-human epidermal growth factor receptor 2 (anti-HER2) nanobody from a naïve llama library by direct panning on whole cells and expressed it fused to Fc and green fluorescent protein...
October 18, 2017: Veterinary and Comparative Oncology
https://www.readbyqxmd.com/read/29045395/single-molecule-imaging-reveals-receptor-g-protein-interactions-at-cell-surface-hot-spots
#11
Titiwat Sungkaworn, Marie-Lise Jobin, Krzysztof Burnecki, Aleksander Weron, Martin J Lohse, Davide Calebiro
G-protein-coupled receptors mediate the biological effects of many hormones and neurotransmitters and are important pharmacological targets. They transmit their signals to the cell interior by interacting with G proteins. However, it is unclear how receptors and G proteins meet, interact and couple. Here we analyse the concerted motion of G-protein-coupled receptors and G proteins on the plasma membrane and provide a quantitative model that reveals the key factors that underlie the high spatiotemporal complexity of their interactions...
October 26, 2017: Nature
https://www.readbyqxmd.com/read/29041922/institute-collection-and-analysis-of-nanobodies-ican-a-comprehensive-database-and-analysis-platform-for-nanobodies
#12
Jing Zuo, Jian Li, Rongxin Zhang, Longsheng Xu, Hanhan Chen, Xiaohuan Jia, Zhipeng Su, Linhong Zhao, Xing Huang, Wei Xie
BACKGROUND: Nanobodies are single-domain antibodies that contain the unique structural and functional properties of naturally-occurring heavy chain in camelidae. As a novel class of antibody, they show many advantages compared with traditional antibodies such as smaller size, higher stability, improved specificity, more easily expressed in microorganisms. These unusual hallmarks make them as promising tools in basic research and clinical practice. Although thousands of nanobodies are known to be published, no single database provides searchable, unified annotation and integrative analysis tools for these various nanobodies...
October 17, 2017: BMC Genomics
https://www.readbyqxmd.com/read/29039819/single-domain-antibodies-as-new-biomarker-detectors
#13
REVIEW
Chiuan Herng Leow, Katja Fischer, Chiuan Yee Leow, Qin Cheng, Candy Chuah, James McCarthy
Biomarkers are defined as indicators of biological processes, pathogenic processes, or pharmacological responses to a therapeutic intervention. Biomarkers have been widely used for early detection, prediction of response after treatment, and for monitoring the progression of diseases. Antibodies represent promising tools for recognition of biomarkers, and are widely deployed as analytical tools in clinical settings. For immunodiagnostics, antibodies are now exploited as binders for antigens of interest across a range of platforms...
October 17, 2017: Diagnostics
https://www.readbyqxmd.com/read/29022874/uncoupling-of-dynamin-polymerization-and-gtpase-activity-revealed-by-the-conformation-specific-nanobody-dynab
#14
Valentina Galli, Rafael Sebastian, Sandrine Moutel, Jason Ecard, Franck Perez, Aurélien Roux
Dynamin is a large GTPase that forms a helical collar at the neck of endocytic pits, and catalyzes membrane fission (Schmid and Frolov, 2011; Ferguson and De Camilli, 2012). Dynamin fission reaction is strictly dependent on GTP hydrolysis, but how fission is mediated is still debated (Antonny et al., 2016): GTP energy could be spent in membrane constriction required for fission, or in disassembly of the dynamin polymer to trigger fission. To follow dynamin GTP hydrolysis at endocytic pits, we generated a conformation-specific nanobody called dynab, that binds preferentially to the GTP hydrolytic state of dynamin-1...
October 12, 2017: ELife
https://www.readbyqxmd.com/read/29017919/whole-cell-biopanning-with-a-synthetic-phage-display-library-of-nanobodies-enabled-the-recovery-of-follicle-stimulating-hormone-receptor-inhibitors
#15
Ronan Crepin, Gianluca Veggiani, Selma Djender, Anne Beugnet, François Planeix, Christophe Pichon, Sandrine Moutel, Sebastian Amigorena, Franck Perez, Nicolae Ghinea, Ario de Marco
Antibodies are essential reagents that are increasingly used in diagnostics and therapy. Their specificity and capacity to recognize their native antigen are critical characteristics for their in vivo application. Follicle-stimulating hormone receptor is a GPCR protein regulating ovarian follicular maturation and spermatogenesis. Recently, its potentiality as a cancer biomarker has been demonstrated but no antibody suitable for in vivo tumor targeting and treatment has been characterized so far. In this paper we describe the first successful attempt to recover recombinant antibodies against the FSHR and that: i) are directly panned from a pre-immune library using whole cells expressing the target receptor at their surface; ii) show inhibitory activity towards the FSH-induced cAMP accumulation; iii) do not share the same epitope with the natural binder FSH; iv) can be produced inexpensively as mono- or bivalent functional molecules in the bacterial cytoplasm...
December 2, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28994409/introducing-site-specific-cysteines-into-nanobodies-for-mercury-labelling-allows-de-novo-phasing-of-their-crystal-structures
#16
Simon Boje Hansen, Nick Stub Laursen, Gregers Rom Andersen, Kasper R Andersen
The generation of high-quality protein crystals and the loss of phase information during an X-ray crystallography diffraction experiment represent the major bottlenecks in the determination of novel protein structures. A generic method for introducing Hg atoms into any crystal independent of the presence of free cysteines in the target protein could considerably facilitate the process of obtaining unbiased experimental phases. Nanobodies (single-domain antibodies) have recently been shown to promote the crystallization and structure determination of flexible proteins and complexes...
October 1, 2017: Acta Crystallographica. Section D, Structural Biology
https://www.readbyqxmd.com/read/28960485/engineered-ferritin-for-magnetogenetic-manipulation-of-proteins-and-organelles-inside-living-cells
#17
Domenik Liße, Cornelia Monzel, Chiara Vicario, John Manzi, Isabelle Maurin, Mathieu Coppey, Jacob Piehler, Maxime Dahan
Magnetogenetics is emerging as a novel approach for remote-controlled manipulation of cellular functions in tissues and organisms with high spatial and temporal resolution. A critical, still challenging issue for these techniques is to conjugate target proteins with magnetic probes that can satisfy multiple colloidal and biofunctional constraints. Here, semisynthetic magnetic nanoparticles are tailored based on human ferritin coupled to monomeric enhanced green fluorescent protein (mEGFP) for magnetic manipulation of proteins inside living cells...
September 28, 2017: Advanced Materials
https://www.readbyqxmd.com/read/28955041/erratum-identification-of-a-nanobody-specific-to-a-human-pulmonary-surfactant-protein-a
#18
Xian He, Shan-Mei Wang, Zhao Fang Yin, Meng-Meng Zhao, Nan Li, Feng Yu, Liu-Sheng Wang, Yang Hu, Yu-Kui Du, Shan-Shan Du, Yan Li, Ya-Ru Wei, Shan-Shan Chen, Jian-Hua He, Dong Weng, Hui-Ping Li
A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.
September 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28938008/vca-nanobodies-target-n-wasp-to-reduce-invadopodium-formation-and-functioning
#19
Tim Hebbrecht, Isabel Van Audenhove, Olivier Zwaenepoel, Adriaan Verhelle, Jan Gettemans
Invasive cancer cells develop small actin-based protrusions called invadopodia, which perform a primordial role in metastasis and extracellular matrix remodelling. Neural Wiskott-Aldrich syndrome protein (N-WASp) is a scaffold protein which can directly bind to actin monomers and Arp2/3 and is a crucial player in the formation of an invadopodium precursor. Expression modulation has pointed to an important role for N-WASp in invadopodium formation but the role of its C-terminal VCA domain in this process remains unknown...
2017: PloS One
https://www.readbyqxmd.com/read/28915239/structural-basis-for-the-high-specificity-of-a-trypanosoma-congolense-immunoassay-targeting-glycosomal-aldolase
#20
Joar Pinto, Steven Odongo, Felicity Lee, Vaiva Gaspariunaite, Serge Muyldermans, Stefan Magez, Yann G-J Sterckx
BACKGROUND: Animal African trypanosomosis (AAT) is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a Nanobody (Nb474) that was employed to develop a homologous sandwich ELISA targeting T. congolense fructose-1,6-bisphosphate aldolase (TcoALD). Despite the high sequence identity between trypanosomatid aldolases, the Nb474-based immunoassay is highly specific for T...
September 2017: PLoS Neglected Tropical Diseases
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