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https://www.readbyqxmd.com/read/28316235/evaluation-of-nanobody-conjugates-and-protein-fusions-as-bioanalytical-reagents
#1
Virginia J Bruce, Brian R McNaughton
Enzyme-linked immunosorbent assay (ELISA), flow cytometry, and Western blot are common bioanalytical techniques. Successful execution traditionally requires the use of one or more commercially available antibody-small-molecule dye, or antibody-reporter protein conjugates that recognize relatively short peptide tags (<15 amino acids). However, the size of antibodies, and their molecular complexity (by virtue of post-translational disulfide formation and glycosylation) typically requires either expression in mammalian cells or purification from immunized mammals...
March 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28303121/identification-of-lama-glama-as-reservoirs-for-acinetobacter-lwoffii
#2
Martín M Ledesma, Ailén M Díaz, Claudia Barberis, Carlos Vay, Marcela A Manghi, Juliana Leoni, Marisa S Castro, Alejandro Ferrari
South American Camelids have an increasing relevance in local economies, worldwide. These animals are bred for their meat, fur and as companion and therapy animals. Thus, their sanitary status should be well-established. According to the OIE (World Organization for Animal Health), respiratory infections mainly produced by Pasteurella spp. have been reported for camelids. It has been stated that this microorganism causes a mild disease, although many authors report it is an important cause of mortality among alpacas...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28290309/polydactyly-suggesting-local-husbandry-of-pre-columbian-camelids-a-case-from-castillo-de-huarmey-archaeological-site-northern-coast-of-peru
#3
Weronika Tomczyk, Miłosz Giersz
Three camelid metapodials with polydactyly (additional digits) were found at the Wari culture archaeological site (dated to the Middle Horizon) of Castillo de Huarmey. The anomalous bones were excavated among numerous remains, and presumably represent animals that were sacrificed within the principal mortuary mausoleum. The bones derive from at least two individuals. The etiology of the deformities remains unknown, but the most probable causes include low genetic diversity in the herd or unintended effect of selective breeding...
March 2017: International Journal of Paleopathology
https://www.readbyqxmd.com/read/28285402/a-semi-empirical-glycosylation-model-of-a-camelid-monoclonal-antibody-under-hypothermia-cell-culture-conditions
#4
Hengameh Aghamohseni, Maureen Spearman, Kaveh Ohadi, Katrin Braasch, Murray Moo-Young, Michael Butler, Hector M Budman
The impact of cell culture environment on the glycan distribution of a monoclonal antibody (mAb) has been investigated through a combination of experiments and modeling. A newly developed CHO DUXB cell line was cultivated at two levels of initial Glutamine (Gln) concentrations (0, 4 mM) and incubation temperatures of (33 and 37 °C) in batch operation mode. Hypothermia was applied either through the entire culture duration or only during the post-exponential phase. Beyond reducing cell growth and increasing productivity, hypothermia significantly altered the galactosylation index profiles as compared to control conditions...
March 11, 2017: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28239762/-1-h-13-c-and-15-n-backbone-chemical-shift-assignments-of-camelid-single-domain-antibodies-against-active-state-%C3%A2%C2%B5-opioid-receptor
#5
Remy Sounier, Yinshan Yang, Joanna Hagelberger, Sébastien Granier, Hélène Déméné
Nanobodies are single chain antibodies that have become a highly valuable and versatile tool for biomolecular and therapeutic research. One application field is the stabilization of active states of flexible proteins, among which G-protein coupled receptors represent a very important class of membrane proteins. Here we present the backbone and side-chain assignment of the (1)H, (13)C and (15)N resonances of Nb33 and Nb39, two nanobodies that recognize and stabilize the µ-opioid receptor to opioids in its active agonist-bound conformation...
February 26, 2017: Biomolecular NMR Assignments
https://www.readbyqxmd.com/read/28214589/enhanced-expression-and-purification-of-camelid-single-domain-vhh-antibodies-from-classical-inclusion-bodies
#6
Maristella Maggi, Claudia Scotti
Single domain antibodies (sdAbs) are small antigen-binding domains derived from naturally occurring, heavy chain-only immunoglobulins isolated from camelid and sharks. They maintain the same binding capability of full-length IgGs but with improved thermal stability and permeability, which justifies their scientific, medical and industrial interest. Several described recombinant forms of sdAbs have been produced in different hosts and with different strategies. Here we present an optimized method for a time-saving, high yield production and extraction of a poly-histidine-tagged sdAb from Escherichia coli classical inclusion bodies...
February 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28209904/noninvasive-imaging-of-human-immune-responses-in-a-human-xenograft-model-of-graft-versus-host-disease
#7
Catharina H M J Van Elssen, Mohammad Rashidian, Vladimir Vrbanac, Kai W Wucherpfennig, Zeina El Habre, Jana Sticht, Christian Freund, Johanne Jacobsen, Juanjo Cragnolini, Jessica Ingram, Loes Plaisier, Eric Spierings, Andrew M Tager, Hidde Ploegh
The immune system plays a crucial role in many diseases. Activation or suppression of immunity is often related to clinical outcome. Methods to explore the dynamics of immune responses are important to elucidate their role in conditions characterized by inflammation, such as infectious disease, cancer or auto-immunity. Immuno-PET is a non-invasive method by which disease and immune cell infiltration can be explored simultaneously. Using radiolabeled antibodies or fragments derived from them, it is possible to image disease-specific antigens and immune cell subsets...
February 16, 2017: Journal of Nuclear Medicine: Official Publication, Society of Nuclear Medicine
https://www.readbyqxmd.com/read/28190501/middle-east-respiratory-syndrome-mers-coronavirus-and-dromedaries
#8
REVIEW
Ulrich Wernery, Susanna K P Lau, Patrick C Y Woo
Middle East Respiratory Syndrome (MERS) is a zoonotic viral disease that can be transmitted from dromedaries to human beings. More than 1500 cases of MERS have been reported in human beings to date. Although MERS has been associated with 30% case fatality in human beings, MERS coronavirus (MERS-CoV) infection in dromedaries is usually asymptomatic. In rare cases, dromedaries may develop mild respiratory signs. No MERS-CoV or antibodies against the virus have been detected in camelids other than dromedaries...
February 2017: Veterinary Journal
https://www.readbyqxmd.com/read/28181310/localization-of-orexin-b-and-receptor-2-for-orexins-in-testicular-cytotypes-of-the-camelid-alpaca-vicugna-pacos
#9
G Liguori, C Squillacioti, L Assisi, N Mirabella, E Langella, A Costagliola, A Vittoria
The orexins A (OxA) and B (OxB) are two hypothalamic peptides involved in many physiological functions of the mammalian body. They act through the binding of two G-coupled receptors named receptor 1 (OX1 ) and receptor 2 (OX2 ) for orexins. The first receptor is specific for OxA, while the second binds both the substances with equal affinity. The orexins and the relative receptors have been traced by means of different techniques also at the periphery of the body and particularly in the adrenals, and in gastrointestinal and genital organs...
February 8, 2017: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/28166983/maternal-recognition-of-pregnancy-and-implantation-are-not-associated-with-an-interferon-response-of-the-endometrium-to-the-presence-of-the-conceptus-in-dromedary-camel
#10
Ahmed Sabry Salah Abdoon, Corrine Giraud-Delville, Omaima Mohamed Kandil, Annelye Kerboeuf-Giraud, Caroline Eozénou, Anais Vitorino Carvalho, Skidmore Julian, Olivier Sandra
Maternal recognition of pregnancy (MRP) and implantation involve appropriate interactions between the elongating conceptus and the receptive endometrium that will condition development of the feto-placental unit to term. Molecular mechanisms that take place at the conceptus-endometrium interface during early pregnancy have been extensively investigated in domestic ungulates but they are still poorly understood in camelids including the dromedary camel (Camelus dromedarius), a domestic species with important economic and social roles in arid and semi-arid areas...
March 1, 2017: Theriogenology
https://www.readbyqxmd.com/read/28166931/evaluation-of-a-panel-of-antibodies-for-the-immunohistochemical-identification-of-immune-cells-in-paraffin-embedded-lymphoid-tissues-of-new-and-old-world-camelids
#11
Ann-Kathrin Uhde, Annika Lehmbecker, Wolfgang Baumgärtner, Ingo Spitzbarth
Different species of camelids play an important role in the epidemiology of various emerging infectious diseases such as Middle East respiratory syndrome. For precise investigations of the immunopathogenesis in these host species, appropriate immunohistochemical markers are highly needed in order to phenotype distinct immune cells populations in camelids. So far, specific immunohistochemical markers for camelid immune cells are rarely commercially available, and cross-reactivity studies are restricted to the use of frozen dromedary tissues...
February 2017: Veterinary Immunology and Immunopathology
https://www.readbyqxmd.com/read/28153778/accelerating-the-clinical-development-of-protein-based-vaccines-for-malaria-by-efficient-purification-using-a-four-amino-acid-c-terminal-c-tag
#12
Jing Jin, Kathryn A Hjerrild, Sarah E Silk, Rebecca E Brown, Geneviève M Labbé, Jennifer M Marshall, Katherine E Wright, Sandra Bezemer, Stine B Clemmensen, Sumi Biswas, Yuanyuan Li, Aadil El-Turabi, Alexander D Douglas, Pim Hermans, Frank J Detmers, Willem A de Jongh, Matthew K Higgins, Rebecca Ashfield, Simon J Draper
Development of bespoke biomanufacturing processes remains a critical bottleneck for translational studies, in particular when modest quantities of a novel product are required for proof-of-concept Phase I/II clinical trials. In these instances the ability to develop a biomanufacturing process quickly and relatively cheaply, without risk to product quality or safety, provides a great advantage by allowing new antigens or concepts in immunogen design to more rapidly enter human testing. These challenges with production and purification are particularly apparent when developing recombinant protein-based vaccines for difficult parasitic diseases, with Plasmodium falciparum malaria being a prime example...
January 30, 2017: International Journal for Parasitology
https://www.readbyqxmd.com/read/28140644/serologic-survey-for-antibodies-against-three-genotypes-of-bovine-parainfluenza-3-virus-in-unvaccinated-ungulates-in-alabama
#13
Benjamin W Newcomer, John D Neill, Patricia K Galik, Kay P Riddell, Yijing Zhang, Thomas Passler, Binu T Velayudhan, Paul H Walz
OBJECTIVE To determine titers of serum antibodies against 3 genotypes of bovine parainfluenza 3 virus (BPI3V) in unvaccinated ungulates in Alabama. ANIMALS 62 cattle, goats, and New World camelids from 5 distinct herds and 21 captured white-tailed deer. PROCEDURES Serum samples were obtained from all animals for determination of anti-BPI3V antibody titers, which were measured by virus neutralization assays that used indicator (reference) viruses from each of the 3 BPI3V genotypes (BPI3V-A, BPI3V-B, and BPI3V-C)...
February 2017: American Journal of Veterinary Research
https://www.readbyqxmd.com/read/28134246/discovery-of-a-junctional-epitope-antibody-that-stabilizes-il-6-and-gp80-protein-protein-interaction-and-modulates-its-downstream-signaling
#14
Ralph Adams, Rebecca J Burnley, Chiara R Valenzano, Omar Qureshi, Carl Doyle, Simon Lumb, Maria Del Carmen Lopez, Robert Griffin, David McMillan, Richard D Taylor, Chris Meier, Prashant Mori, Laura M Griffin, Ulrich Wernery, Jörg Kinne, Stephen Rapecki, Terry S Baker, Alastair D G Lawson, Michael Wright, Anna Ettorre
Protein:protein interactions are fundamental in living organism homeostasis. Here we introduce VHH6, a junctional epitope antibody capable of specifically recognizing a neo-epitope when two proteins interact, albeit transiently, to form a complex. Orthogonal biophysical techniques have been used to prove the "junctional epitope" nature of VHH6, a camelid single domain antibody recognizing the IL-6-gp80 complex but not the individual components alone. X-ray crystallography, HDX-MS and SPR analysis confirmed that the CDR regions of VHH6 interact simultaneously with IL-6 and gp80, locking the two proteins together...
January 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28131818/improved-production-of-single-domain-antibodies-with-two-disulfide-bonds-by-co-expression-of-chaperone-proteins-in-the-escherichia-coli-periplasm
#15
Lisa C Shriver-Lake, Ellen R Goldman, Daniel Zabetakis, George P Anderson
Single domain antibodies are recombinantly expressed variable domains derived from camelid heavy chain antibodies. Natural single domain antibodies can have noncanonical disulfide bonds between their complementarity-determining regions that help position the binding site. In addition, engineering a second disulfide bond serves to tie together β-sheets thereby inhibiting unfolding. Unfortunately, the additional disulfide bond often significantly decreases yield, presumably due to formation of incorrect disulfide bonds during the folding process...
January 26, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28114943/joining-the-in-vitro-immunization-of-alpaca-lymphocytes-and-phage-display-rapid-and-cost-effective-pipeline-for-sdab-synthesis
#16
Lubos Comor, Saskia Dolinska, Katarina Bhide, Lucia Pulzova, Irene Jiménez-Munguía, Elena Bencurova, Zuzana Flachbartova, Lenka Potocnakova, Evelina Kanova, Mangesh Bhide
BACKGROUND: Camelids possess unique functional heavy chain antibodies, which can be produced and modified in vitro as a single domain antibody (sdAb or nanobody) with full antigen binding ability. Production of sdAb in conventional manner requires active immunization of Camelidae animal, which is laborious, time consuming, costly and in many cases not feasible (e.g. in case of highly toxic or infectious antigens). RESULTS: In this study, we describe an alternative pipeline that includes in vitro stimulation of naïve alpaca B-lymphocytes by antigen of interest (in this case endothelial cell binding domain of OspA of Borrelia) in the presence of recombinant alpaca interleukins 2 and 4, construction of sdAb phage library, selection of antigen specific sdAb expressed on phages (biopanning) and confirmation of binding ability of sdAb to the antigen...
January 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28065273/screening-and-characterization-strategies-for-nanobodies-targeting-membrane-proteins
#17
S Veugelen, M Dewilde, B De Strooper, L Chávez-Gutiérrez
The study of membrane protein function and structure requires their successful detection, expression, solubilization, and/or reconstitution, which poses a challenging task and relies on the availability of suitable tools. Several research groups have successfully applied Nanobodies in the purification, as well as the functional and structural characterization of membrane proteins. Nanobodies are small, single-chain antibody fragments originating from camelids presenting on average a longer CDR3 which enables them to bind in cavities and clefts (such as active and allosteric sites)...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28065171/zoonotic-diseases-in-south-american-camelids-in-england-and-wales
#18
K Halsby, D F Twomey, C Featherstone, A Foster, A Walsh, K Hewitt, D Morgan
The number of South American camelids (SACs) in England and Wales is increasing and with this comes a risk of new and emerging infections. Although classified as livestock, these animals are also treated as pets and may be in regular contact with humans. This paper reviews zoonotic diseases that have been identified in SACs in England and Wales, and which pose a potential risk to human health. We also highlight the importance of surveillance continuing to capture information on infections in SACs for the protection of both public and animal health...
January 9, 2017: Epidemiology and Infection
https://www.readbyqxmd.com/read/28043352/current-status-and-future-direction-of-cryopreservation-of-camelid-embryos
#19
REVIEW
M Herrid, G Vajta, J A Skidmore
Over the past 3 decades, and similar to the horse industry, fresh embryo transfer has been widely practiced on large commercial scales in different camelid species, especially the dromedary camel and alpaca. However, the inability to cryopreserve embryos significantly reduces its broader application, and as such limits the capacity to utilize elite genetic resources internationally. In addition, cryopreservation of the semen of camelids is also difficult, suggesting an extreme sensitivity of the germplasm to cooling and freezing...
February 2017: Theriogenology
https://www.readbyqxmd.com/read/28012779/vaccination-of-alpacas-against-rift-valley-fever-virus-safety-immunogenicity-and-pathogenicity-of-mp-12-vaccine
#20
M Rissmann, R Ulrich, C Schröder, B Hammerschmidt, D Hanke, C Mroz, M H Groschup, M Eiden
Rift Valley fever (RVF) is an emerging zoonosis of major public health concern in Africa and Arabia. Previous outbreaks attributed camelids a significant role in the epidemiology of Rift Valley fever virus (RVFV), making them an important target species for vaccination. Using three alpacas as model-organisms for dromedary camels, the safety, immunogenicity and pathogenicity of the MP-12 vaccine were evaluated in this study. To compare both acute and subacute effects, animals were euthanized at 3 and 31days post infection (dpi)...
December 21, 2016: Vaccine
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