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https://www.readbyqxmd.com/read/27929050/first-complete-mitochondrial-genome-data-from-ancient-south-american-camelids-the-mystery-of-the-chilihueques-from-isla-mocha-chile
#1
Michael Westbury, Stefan Prost, Andrea Seelenfreund, José-Miguel Ramírez, Elizabeth A Matisoo-Smith, Michael Knapp
In South American societies, domesticated camelids were of great cultural importance and subject to trade and translocation. South American camelids were even found on remote and hard to reach islands, emphasizing their importance to historic and pre-historic South American populations. Isla Mocha, a volcanic island 35 km offshore of Central-South Chile, is an example of such an island. When Dutch and Spanish explorers reached the island in the early 17th century, they found that domesticated camelids called "chilihueque" played a major role in the island's society...
December 8, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27905655/-gastrointestinal-bezoars
#2
Ricardo Espinoza González
Gastrointestinal bezoars are a concretion of indigested material that can be found in the gastrointestinal tract of humans and some animals. This material forms an intraluminal mass, more commonly located in the stomach. During a large period of history animal bezoars were considered antidotes to poisons and diseases. We report a historical overview since bezoars stones were thought to have medicinal properties. This magic conception was introduced in South America by Spanish conquerors. In Chile, bezoars are commonly found in a camelid named guanaco (Lama guanicoe)...
August 2016: Revista Médica de Chile
https://www.readbyqxmd.com/read/27903650/structural-analysis-of-single-domain-antibodies-bound-to-a-second-neutralizing-hot-spot-on-ricin-toxin-s-enzymatic-subunit
#3
Michael J Rudolph, David J Vance, Michael S Cassidy, Yinghui Rong, Nicholas J Mantis
Ricin toxin is a heterodimer consisting of RTA, a ribosome-inactivating protein, and RTB, a lectin that facilitates receptor-mediated uptake into mammalian cells. In previous studies, we demonstrated that toxin-neutralizing antibodies target four spatially distinct hotspots on RTA, which we refer to as epitope Clusters I-IV. In this report we identified and characterized three single domain camelid antibodies (VHH) against Cluster II. One of these VHHs, V5E1, ranks as one of the most potent ricin-neutralizing antibodies described to date...
November 30, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27881685/generation-of-camelid-vhh-bispecific-constructs-via-in-cell-intein-mediated-protein-trans-splicing
#4
Yuki Shibuya, Natsuki Haga, Ryutaro Asano, Hikaru Nakazawa, Takamitsu Hattori, Daisuke Takeda, Aruto Sugiyama, Reiko Kurotani, Izumi Kumagai, Mitsuo Umetsu, Koki Makabe
Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity...
November 23, 2016: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/27865111/camelid-and-shark-single-domain-antibodies-structural-features-and-therapeutic-potential
#5
REVIEW
Doreen Könning, Stefan Zielonka, Julius Grzeschik, Martin Empting, Bernhard Valldorf, Simon Krah, Christian Schröter, Carolin Sellmann, Björn Hock, Harald Kolmar
In addition to canonical antibodies composed of heavy and light chains, the adaptive immune systems of camelids and cartilaginous fish comprise heavy-chain only isotypes (HcAb) devoid of light chains, where antigen-binding is mediated exclusively by one variable domain. Due to their inherent favorable attributes, such as high affinity and specificity for their cognate antigen, extraordinary stability, small size and, most importantly, the possibility to complement classical antibodies in terms of 'drugable' target-space, HcAb-derived entities evolved as promising candidates for biomedical applications of which many have already proven to be successful in early stage clinical trials...
November 16, 2016: Current Opinion in Structural Biology
https://www.readbyqxmd.com/read/27862476/analysis-of-the-binding-loops-configuration-and-surface-adaptation-of-different-crystallized-single-domain-antibodies-in-response-to-various-antigens
#6
Mohammed M Al Qaraghuli, Valerie A Ferro
Monoclonal antibodies have revolutionized the biomedical field through their ubiquitous utilization in different diagnostics and therapeutic applications. Despite this widespread use, their large size and structural complexity have limited their versatility in specific applications. The antibody variable region that is responsible for binding antigen is embodied within domains that can be rescued individually as single-domain antibody (sdAb) fragments. Because of the unique characteristics of sdAbs, such as low molecular weight, high physicochemical stability, and the ability to bind antigens inaccessible to conventional antibodies, they represent a viable alternative to full-length antibodies...
November 16, 2016: Journal of Molecular Recognition: JMR
https://www.readbyqxmd.com/read/27862386/no-detectable-carotenoid-concentrations-in-serum-of-llamas-and-alpacas
#7
J Raila, F J Schweigert, A Stanitznig, B Lambacher, S Franz, S Baldermann, T Wittek
Carotenoids are lipid-soluble pigments and important for a variety of physiological functions. They are major dietary vitamin A precursors and act as lipophilic antioxidants in a variety of tissues and are associated with important health benefits in humans and animals. All animals must acquire carotenoids from their diet, but to our knowledge, there are no studies investigating the intestinal carotenoid absorption and their blood concentrations in New World camelids. The present study aimed to assess the serum concentrations of selected carotenoids in llamas (n = 13) and alpacas (n = 27)...
November 9, 2016: Journal of Animal Physiology and Animal Nutrition
https://www.readbyqxmd.com/read/27852863/cryoem-structures-of-expanded-poliovirus-with-vhhs-sample-the-conformational-repertoire-of-the-expanded-state
#8
Mike Strauss, Lise Schotte, Krishanthi S Karunatilaka, David J Filman, James M Hogle
: Using cryoelectron microscopy, expanded 80S-like poliovirions were visualized in complexes with four 80S-specific camelid VHHs (Nanobodies®). In all four complexes, the VHHs bind to a site on the top surface of capsid protein VP3, which is hidden in native virus. Interestingly, although the four VHHs bind to the same site, the structures of the expanded virus differ in detail in each complex, suggesting that each of the nanobodies has sampled a range of low energy structures available to the expanded virion...
November 16, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27816596/rapid-capture-and-labeling-of-cells-on-single-domain-antibodies-functionalized-flow-cell
#9
Guan-Yu Chen, Zeyang Li, Joao N Duarte, Alexandre Esteban, Ross W Cheloha, Christopher S Theile, Gerald R Fink, Hidde L Ploegh
Current techniques to characterize leukocyte subgroups in blood require long sample preparation times and sizable sample volumes. A simplified method for leukocyte characterization using smaller blood volumes would thus be useful in diagnostic settings. Here we describe a flow system comprised of two functionalized graphene oxide (GO) surfaces that allow the capture of distinct leukocyte populations from small volumes blood using camelid single-domain antibodyfragments (VHHs) as capture agents. We used site-specifically labeled leukocytes to detect and identify cells exposed to fungal challenge...
October 5, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27805447/evaluation-of-two-platelet-rich-plasma-processing-methods-and-two-platelet-activation-techniques-for-use-in-llamas-and-alpacas
#10
Stacy A Semevolos, Cori D Youngblood, Stephanie K Grissom, M Elena Gorman, Maureen K Larson
OBJECTIVE To evaluate 2 processing methods (commercial kit vs conical tube centrifugation) for preparing platelet rich plasma (PRP) for use in llamas and alpacas. SAMPLES Blood samples (30 mL each) aseptically collected from 6 healthy llamas and 6 healthy alpacas. PROCEDURES PRP was prepared from blood samples by use of a commercial kit and by double-step conical tube centrifugation. A CBC was performed for blood and PRP samples. Platelets in PRP samples were activated by means of a freeze-thaw method with or without 23mM CaCl2, and concentrations of platelet-derived growth factor-BB and transforming growth factor-β1 were measured...
November 2016: American Journal of Veterinary Research
https://www.readbyqxmd.com/read/27791182/crystal-structure-of-a-lacy-nanobody-complex-in-a-periplasmic-open-conformation
#11
Xin Jiang, Irina Smirnova, Vladimir Kasho, Jianping Wu, Kunio Hirata, Meng Ke, Els Pardon, Jan Steyaert, Nieng Yan, H Ronald Kaback
The lactose permease of Escherichia coli (LacY), a dynamic polytopic membrane protein, catalyzes galactoside-H(+) symport and operates by an alternating access mechanism that exhibits multiple conformations, the distribution of which is altered by sugar binding. We have developed single-domain camelid nanobodies (Nbs) against a mutant in an outward (periplasmic)-open conformation to stabilize this state of the protein. Here we describe an X-ray crystal structure of a complex between a double-Trp mutant (Gly46→Trp/Gly262→Trp) and an Nb in which free access to the sugar-binding site from the periplasmic cavity is observed...
November 1, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27762054/ovulation-inducing-factor-oif-ngf-in-seminal-plasma-a-review-and-update
#12
G P Adams, M H Ratto, M E Silva, R A Carrasco
The ovulation-inducing effect of seminal plasma was first reported in Bactrian camels over 30 years ago, and the entity responsible was dubbed 'ovulation-inducing factor' (OIF). More recent studies, primarily in llamas and alpacas, characterized the biological and chemical properties of OIF and ultimately identified it as βNGF. This recent discovery has allowed a convergence of knowledge previously separated by discipline and by mechanism; that is, neurobiology and reproductive biology, and autocrine/paracrine vs endocrine...
October 2016: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/27721040/inhibition-of-glycosylation-on-a-camelid-antibody-uniquely-affects-its-fc%C3%AE-ri-binding-activity
#13
Natalie Krahn, Maureen Spearman, Markus Meier, July Dorion-Thibaudeau, Matthew McDougall, Trushar R Patel, Gregory De Crescenzo, Yves Durocher, Jörg Stetefeld, Michael Butler
Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity is well known. However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. This study analysed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80 kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150 kDa)...
October 6, 2016: European Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/27686537/thermodynamics-of-nanobody-binding-to-lactose-permease
#14
Parameswaran Hariharan, Magnus Andersson, Xiaoxu Jiang, Els Pardon, Jan Steyaert, H Ronald Kaback, Lan Guan
Camelid nanobodies (Nbs) raised against the outward-facing conformer of a double-Trp mutant of the lactose permease of Escherichia coli (LacY) stabilize the permease in outward-facing conformations. Isothermal titration calorimetry is applied herein to dissect the binding thermodynamics of two Nbs, one that markedly improves access to the sugar-binding site and another that dramatically increases the affinity for galactoside. The findings presented here show that both enthalpy and entropy contribute favorably to binding of the Nbs to wild-type (WT) LacY and that binding of Nb to double-Trp mutant G46W/G262W is driven by a greater enthalpy at an entropic penalty...
October 12, 2016: Biochemistry
https://www.readbyqxmd.com/read/27677057/camelid-vh-h-affinity-ligands-enable-separation-of-closely-related-biopharmaceuticals
#15
Timothy M Pabst, Michaela Wendeler, Xiangyang Wang, Sandra Bezemer, Pim Hermans, Alan K Hunter
Interest in new and diverse classes of molecules such as recombinant toxins, enzymes, and blood factors continues to grow for use a biotherapeutics. Compared to monoclonal antibodies, these novel drugs typically lack a commercially available affinity chromatography option, which leads to greater process complexity, longer development timelines, and poor platformability. To date, for both monoclonal antibodies and novel molecules, affinity chromatography has been mostly reserved for separation of process-related impurities such as host cell proteins and DNA...
September 27, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27671875/camelid-single-domain-antibodies-a-versatile-tool-for-in-vivo-imaging-of-extracellular-and-intracellular-brain-targets
#16
Tengfei Li, Matthias Vandesquille, Fani Koukouli, Clémence Dudeffant, Ihsen Youssef, Pascal Lenormand, Christelle Ganneau, Uwe Maskos, Christian Czech, Fiona Grueninger, Charles Duyckaerts, Marc Dhenain, Sylvie Bay, Benoît Delatour, Pierre Lafaye
Detection of intracerebral targets with imaging probes is challenging due to the non-permissive nature of blood-brain barrier (BBB). The present work describes two novel single-domain antibodies (VHHs or nanobodies) that specifically recognize extracellular amyloid deposits and intracellular tau neurofibrillary tangles, the two core lesions of Alzheimer's disease (AD). Following intravenous administration in transgenic mouse models of AD, in vivo real-time two-photon microscopy showed gradual extravasation of the VHHs across the BBB, diffusion in the parenchyma and labeling of amyloid deposits and neurofibrillary tangles...
October 5, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/27670069/the-structure-of-a-furin-antibody-complex-explains-non-competitive-inhibition-by-steric-exclusion-of-substrate-conformers
#17
Sven O Dahms, John W M Creemers, Yvonne Schaub, Gleb P Bourenkov, Thomas Zögg, Hans Brandstetter, Manuel E Than
Proprotein Convertases (PCs) represent highly selective serine proteases that activate their substrates upon proteolytic cleavage. Their inhibition is a promising strategy for the treatment of cancer and infectious diseases. Inhibitory camelid antibodies were developed, targeting the prototypical PC furin. Kinetic analyses of them revealed an enigmatic non-competitive mechanism, affecting the inhibition of large proprotein-like but not small peptidic substrates. Here we present the crystal structures of furin in complex with the antibody Nb14 and of free Nb14 at resolutions of 2...
September 27, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27669966/identification-of-useful-nanobodies-by-phage-display-of-immune-single-domain-libraries-derived-from-camelid-heavy-chain-antibodies
#18
Ema Romão, Francisco Morales-Yánez, Yaozhong Hu, Maxine Crauwels, Pieter De Pauw, Gholamreza Ghassanzadeh Hassanzadeh, Nick Devoogdt, Chloé Ackaert, Cécile Vincke, Serge Muyldermans
BACKGROUND: The discovery of functional heavy chain-only antibodies devoid of light chains in sera of camelids and sharks in the early nineties provided access to the generation of minimal-sized, single-domain, in vivo affinity-matured, recombinant antigen-binding fragments, also known as Nanobodies. METHODS: Recombinant DNA technology and adaptation of phage display vectors form the basis to construct large naïve, synthetic or medium sized immune libraries from where multiple Nanobodies have been retrieved...
September 23, 2016: Current Pharmaceutical Design
https://www.readbyqxmd.com/read/27639582/biochemical-characterization-of-the-detoxifying-enzyme-glutathione-transferase-p1-1-from-the-camel-camelus-dromedarius
#19
Fereniki Perperopoulou, Farid S Ataya, Dalia Fouad, Ajamaluddin Malik, Hesham Mahmoud Saeed, Nikolaos E Labrou
Glutathione transferase (GST, EC 2.5.1.18) is a primary line of defense against toxicities of electrophile compounds and oxidative stress and therefore is involved in stress-response and cell detoxification. In the present study, we investigated the catalytic and structural properties of the glutathione transferase (GST) isoenzyme P1-1 from Camelus dromedarius (CdGSTP1-1). Recombinant CdGSTP1-1 was produced in Escherichia coli BL21(DE3) and purified to electrophoretic homogeneity. Kinetic analysis revealed that CdGSTP1-1 displays broad substrate specificity and shows high activity towards halogenated aryl-compounds, isothiocyanates and hydroperoxides...
September 17, 2016: Cell Biochemistry and Biophysics
https://www.readbyqxmd.com/read/27631624/a-rational-engineering-strategy-for-designing-protein-a-binding-camelid-single-domain-antibodies
#20
Kevin A Henry, Traian Sulea, Henk van Faassen, Greg Hussack, Enrico O Purisima, C Roger MacKenzie, Mehdi Arbabi-Ghahroudi
Staphylococcal protein A (SpA) and streptococcal protein G (SpG) affinity chromatography are the gold standards for purifying monoclonal antibodies (mAbs) in therapeutic applications. However, camelid VHH single-domain Abs (sdAbs or VHHs) are not bound by SpG and only sporadically bound by SpA. Currently, VHHs require affinity tag-based purification, which limits their therapeutic potential and adds considerable complexity and cost to their production. Here we describe a simple and rapid mutagenesis-based approach designed to confer SpA binding upon a priori non-SpA-binding VHHs...
2016: PloS One
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