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https://www.readbyqxmd.com/read/29031230/astatine-211-labeled-anti-her2-5f7-single-domain-antibody-fragment-conjugates-radiolabeling-and-preliminary-evaluation
#1
Jaeyeon Choi, Ganesan Vaidyanathan, Eftychia Koumarianou, Choong Mo Kang, Michael R Zalutsky
INTRODUCTION: Derived from heavy chain only camelid antibodies, ~15-kDa single-domain antibody fragments (sdAbs) are an attractive platform for developing molecularly specific imaging probes and targeted radiotherapeutics. The rapid tumor accumulation and normal tissue clearance of sdAbs might be ideal for use with (211)At, a 7.2-h half-life α-emitter, if appropriate labeling chemistry can be devised to trap (211)At in cancer cells after sdAb binding. This study evaluated two reagents, [(211)At]SAGMB and iso-[(211)At]SAGMB, for this purpose...
September 19, 2017: Nuclear Medicine and Biology
https://www.readbyqxmd.com/read/29024003/molecular-characterization-of-the-llama-fgf5-gene-and-identification-of-putative-loss-of-function-mutations
#2
M S Daverio, L Vidal-Rioja, E N Frank, F Di Rocco
Llama, the most numerous domestic camelid in Argentina, has good fiber-production ability. Although a few genes related to other productive traits have been characterized, the molecular genetic basis of fiber growth control in camelids is still poorly understood. Fibroblast growth factor 5 (FGF5) is a secreted signaling protein that controls hair growth in humans and other mammals. Mutations in the FGF5 gene have been associated with long-hair phenotypes in several species. Here, we sequenced the llama FGF5 gene, which consists of three exons encoding 813 bp...
October 11, 2017: Animal Genetics
https://www.readbyqxmd.com/read/28986243/construction-of-an-expression-vector-that-uses-the-aph-promoter-for-protein-expression-in-corynebacterium-glutamicum
#3
Wei Zhang, Zihao Zhao, Yankun Yang, Xiuxia Liu, Zhonghu Bai
Corynebacterium glutamicum is an attractive host for the production of heterologous proteins despite its traditional use in fermentative production of amino acids. To enhance the expression levels of target genes, the development of useful promoters is required in the construction of expression systems. Here, we developed a new promoter, the aph promoter from aminoglycoside-3'-phosphotransferase gene, and used it to construct monocistronic and bicistronic expression systems that host different ribosome binding site (RBS) sequences...
October 3, 2017: Plasmid
https://www.readbyqxmd.com/read/28982028/effect-of-age-and-castration-on-serum-anti-m%C3%A3-llerian-hormone-concentration-in-male-alpacas
#4
Michela Ciccarelli, Ahmed Tibary, Alexis J Campbell, Alan J Conley
The synthesis of anti-Müllerian Hormone (AMH) by the Sertoli cells in males is crucial for sexual differentiation. There are no studies on AMH in Camelids. The objectives of this research were to 1) compare AMH serum concentrations in prepubertal and adult male alpacas and 2) determine the effect of castration on these concentrations in adult males to provide a validation of a commercial AMH test in alpacas. Serum samples were obtained from 15 prepubertal animals (5 for each age groups of 6, 7 and 8 months) and from 5 adult males (age 5-9 years), pre- and 24 h post-castration...
September 28, 2017: Theriogenology
https://www.readbyqxmd.com/read/28974337/iodoacetyl-functionalized-pullulan-a-supplemental-enhancer-for-single-domain-antibody-polyclonal-antibody-sandwich-enzyme-linked-immunosorbent-assay-for-detection-of-survivin
#5
Takahiko Matsushita, Hidenao Arai, Tetsuo Koyama, Ken Hatano, Naoto Nemoto, Koji Matsuoka
Survivin, an inhibitor of the apoptosis protein family, is a potent tumor marker for diagnosis and prognosis. The enzyme-linked immunosorbent assay (ELISA) is one of the methods that has been used for detection of survivin. However, ELISA has several disadvantages caused by the use of conventional antibodies, and we have therefore been trying to develop a novel ELISA system using camelid single-domain antibodies (VHHs) as advantageous replacements. Here we report a supplemental approach to improve the VHH-polyclonal antibody sandwich ELISA for survivin detection...
September 23, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28961345/prediction-of-packed-cell-volume-after-whole-blood-transfusion-in-small-ruminants-and-south-american-camelids-80-cases-2006-2016
#6
D Luethy, D Stefanovski, R Salber, R W Sweeney
BACKGROUND: Calculation of desired whole blood transfusion volume relies on an estimate of an animal's circulating blood volume, generally accepted to be 0.08 L/kg or 8% of the animal's body weight in kilograms. OBJECTIVE: To use packed cell volume before and after whole blood transfusion to evaluate the accuracy of a commonly used equation to predict packed cell volume after transfusion in small ruminants and South American camelids; to determine the nature and frequency of adverse transfusion reactions in small ruminants and camelids after whole blood transfusion...
September 29, 2017: Journal of Veterinary Internal Medicine
https://www.readbyqxmd.com/read/28935898/pd-l1-is-an-activation-independent-marker-of-brown-adipocytes
#7
Jessica R Ingram, Michael Dougan, Mohammad Rashidian, Marko Knoll, Edmund J Keliher, Sarah Garrett, Scott Garforth, Olga S Blomberg, Camilo Espinosa, Atul Bhan, Steven C Almo, Ralph Weissleder, Harvey Lodish, Stephanie K Dougan, Hidde L Ploegh
Programmed death ligand 1 (PD-L1) is expressed on a number of immune and cancer cells, where it can downregulate antitumor immune responses. Its expression has been linked to metabolic changes in these cells. Here we develop a radiolabeled camelid single-domain antibody (anti-PD-L1 VHH) to track PD-L1 expression by immuno-positron emission tomography (PET). PET-CT imaging shows a robust and specific PD-L1 signal in brown adipose tissue (BAT). We confirm expression of PD-L1 on brown adipocytes and demonstrate that signal intensity does not change in response to cold exposure or β-adrenergic activation...
September 21, 2017: Nature Communications
https://www.readbyqxmd.com/read/28917312/first-finding-of-nymphal-stages-of-linguatula-serrata-in-a-south-american-camelid-a-vicu%C3%A3-a-from-peru
#8
Luis A Gomez-Puerta, Joel I Pacheco, José M Angulo-Tisoc, Maria T Lopez-Urbina, Armando E Gonzalez
Linguatula serrata, a pentastomid, was found parasitizing the lungs of a vicuña (Vicugna vicugna) from Cuzco, Peru. A total of 13 larvae were found encysted in the parenchymal tissue of the lungs. All larvae were identified as nymphal stages of L. serrata by morphological methods Diagnosis was confirmed by molecular analysis amplifying the cytochrome c oxidase 1 gene of three nymphs. Nucleotide sequences from the isolates were compared to previous sequences from GenBank, and it showed high similarity between them (>99%)...
September 15, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28884482/histological-characterization-of-umbilical-cord-in-alpaca-vicugna-pacos
#9
L M Barrios-Arpi, J-L Rodríguez Gutiérrez, B Lopez-Torres
The histomorphometric features of umbilical cord constituents in seven foetuses of alpaca (Vicugna pacos) from Cerro de Pasco, Department, Peru, were determined. Sections of 2-5 cm of umbilical cord were fixed in 10% neutral buffered formalin and processed for light microscopy. Standard histological slides stained with haematoxylin and eosin, Masson's trichrome and Van Gieson's trichrome were obtained. Histologically, common features of umbilical artery and vein were observed as well as mucous connective tissue, some cell features that compound this tissue constituted by cells presented features of myofibroblasts...
September 7, 2017: Anatomia, Histologia, Embryologia
https://www.readbyqxmd.com/read/28871254/single-domain-antibodies-as-versatile-affinity-reagents-for-analytical-and-diagnostic-applications
#10
REVIEW
Gualberto Gonzalez-Sapienza, Martín A Rossotti, Sofía Tabares-da Rosa
With just three CDRs in their variable domains, the antigen-binding site of camelid heavy-chain-only antibodies (HcAbs) has a more limited structural diversity than that of conventional antibodies. Even so, this does not seem to limit their specificity and high affinity as HcAbs against a broad range of structurally diverse antigens have been reported. The recombinant form of their variable domain [nanobody (Nb)] has outstanding properties that make Nbs, not just an alternative option to conventional antibodies, but in many cases, these properties allow them to reach analytical or diagnostic performances that cannot be accomplished with conventional antibodies...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28871252/development-and-characterization-of-a-camelid-single-domain-antibody-urease-conjugate-that-targets-vascular-endothelial-growth-factor-receptor-2
#11
Baomin Tian, Wah Yau Wong, Marni D Uger, Pawel Wisniewski, Heman Chao
Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor, which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFRs). In this study, we describe the development and characterization of V21-DOS47, an immunoconjugate that targets VEGFR2. V21-DOS47 is composed of a camelid single domain anti-VEGFR2 antibody (V21) and the enzyme urease...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28871080/genetically-engineered-red-cells-expressing-single-domain-camelid-antibodies-confer-long-term-protection-against-botulinum-neurotoxin
#12
Nai-Jia Huang, Novalia Pishesha, Jean Mukherjee, Sicai Zhang, Rhogerry Deshycka, Valentino Sudaryo, Min Dong, Charles B Shoemaker, Harvey F Lodish
A short half-life in the circulation limits the application of therapeutics such as single-domain antibodies (VHHs). We utilize red blood cells to prolong the circulatory half-life of VHHs. Here we present VHHs against botulinum neurotoxin A (BoNT/A) on the surface of red blood cells by expressing chimeric proteins of VHHs with Glycophorin A or Kell. Mice whose red blood cells carry the chimeric proteins exhibit resistance to 10,000 times the lethal dose (LD50) of BoNT/A, and transfusion of these red blood cells into naive mice affords protection for up to 28 days...
September 4, 2017: Nature Communications
https://www.readbyqxmd.com/read/28869728/-application-of-nanobody-in-cancer-treatment
#13
Yue Wu, Xiujing Hao, Min Li
Camelidae can produce a unique antibody that lacks light chain called variable heavy chain domain, also known as nanobodies. This antibody contains only one variable region, with high affinity, high stability, strong tissue penetration, efficient expression. Besides, their toxicity and immunogenicity are both low to be used for both therapeutic and diagnostic applications, as well as research tools. In this review, we discuss how nanobody has been explored as therapeutics in oncology, and provide ideas for the further development of nanobody...
July 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28858207/x-chromosome-evolution-in-cetartiodactyla
#14
Anastasia A Proskuryakova, Anastasia I Kulemzina, Polina L Perelman, Alexey I Makunin, Denis M Larkin, Marta Farré, Anna V Kukekova, Jennifer Lynn Johnson, Natalya A Lemskaya, Violetta R Beklemisheva, Melody E Roelke-Parker, June Bellizzi, Oliver A Ryder, Stephen J O'Brien, Alexander S Graphodatsky
The phenomenon of a remarkable conservation of the X chromosome in eutherian mammals has been first described by Susumu Ohno in 1964. A notable exception is the cetartiodactyl X chromosome, which varies widely in morphology and G-banding pattern between species. It is hypothesized that this sex chromosome has undergone multiple rearrangements that changed the centromere position and the order of syntenic segments over the last 80 million years of Cetartiodactyla speciation. To investigate its evolution we have selected 26 evolutionarily conserved bacterial artificial chromosome (BAC) clones from the cattle CHORI-240 library evenly distributed along the cattle X chromosome...
August 31, 2017: Genes
https://www.readbyqxmd.com/read/28855086/hematocrit-of-mammals-artiodactyla-carnivora-primates-at-1500m-and-2100m-altitudes
#15
Jacopo P Mortola, DeeAnn Wilfong
The rise in hematocrit (Hct) is one of the hallmarks of human acclimatization to high altitude and, in chronic conditions, reflects the hypoxia-induced polycythemia. However, it is not a uniform response among domestic species and it is not found in Andean camelids, species long adapted to high altitudes. Hence, we asked to what extent the polycythemia of humans is common among mammals. Hct data were collected from captive mammals of three orders (Primates, Artiodactyla, Carnivora), 70 specimens of 33 species at ∼1500m altitude (barometric pressure Pb=635mmHg) and 296 specimens of 64 species at ∼2100m (Pb=596mmHg), long-term residents at those altitudes...
July 8, 2017: Zoology: Analysis of Complex Systems, ZACS
https://www.readbyqxmd.com/read/28853185/a-camelid-nanobody-against-egfr-was-easily-obtained-through-refolding-of-inclusion-body-expressed-in-escherichia-coli
#16
Li Xu, Xiaoyu Song, Lingyun Jia
Using anti-EGFR (epidermal growth factor receptor) nanobody is a good choice for diagnoses and therapeutics for high EGFR expression diseases. In the present study, the percentage composition of anti-EGFR nanobody attained 25% of the total cell protein expressed in Escherichia coli BL21 (DE3). However, almost all nanobodies were expressed as inclusion bodies. To acquire active nanobodies, a series of dilution refolding procedures were optimized after inclusion bodies were dissolved into 6 M urea and purified with immobilized metal affinity chromatography...
November 7, 2016: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28848291/trypanosoma-evansi-infection-and-major-risk-factors-for-iranian-one-humped-camels-camelus-dromedarius
#17
Freshteh Mirshekar, Mohammad Yakhchali, Fariborz Shariati-Sharifi
Trypanosoma evansi is a cosmopolitan protozoan which affects camelids and may cause illness and economic losses. The present study was aimed to determine the prevalence and geographic distribution of T. evansi in Iranian one-humped camels of South East Iran. A total of 369 camels were randomly examined from three parts of southeastern Iran from March to September 2015. Blood samples were taken from jugular vein and examined by using thin blood smear and mercuric chloride test. Ninety-five out of 369 examined camels (25...
September 2017: Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
https://www.readbyqxmd.com/read/28830429/molecular-study-on-pasteurella-multocida-and-mannheimia-granulomatis-from-kenyan-camels-camelus-dromedarius
#18
Ilona V Gluecks, Astrid Bethe, Mario Younan, Christa Ewers
BACKGROUND: Outbreaks of a Haemorrhagic Septicaemia (HS) like disease causing large mortalities in camels (Camelus dromedarius) in Asia and in Africa have been reported since 1890. Yet the aetiology of this condition remains elusive. This study is the first to apply state of the art molecular methods to shed light on the nasopharyngeal carrier state of Pasteurellaceae in camels. The study focused on HS causing Pasteurella multocida capsular types B and E. Other Pasteurellaceae, implicated in common respiratory infections of animals, were also investigated...
August 22, 2017: BMC Veterinary Research
https://www.readbyqxmd.com/read/28827559/selection-of-nanobodies-with-broad-neutralizing-potential-against-primary-hiv-1-strains-using-soluble-subtype-c-gp140-envelope-trimers
#19
Kathrin Koch, Sarah Kalusche, Jonathan L Torres, Robyn L Stanfield, Welbeck Danquah, Kamal Khazanehdari, Hagen von Briesen, Eric R Geertsma, Ian A Wilson, Ulrich Wernery, Friedrich Koch-Nolte, Andrew B Ward, Ursula Dietrich
Broadly neutralizing antibodies (bnAbs) against HIV-1 protect from infection and reduce viral load upon therapeutic applications. However no vaccine was able so far to induce bnAbs demanding their expensive biotechnological production. For clinical applications, nanobodies (VHH) derived from heavy chain only antibodies from Camelidae, may be better suited due to their small size, high solubility/stability and extensive homology to human VH3 genes. Here we selected broadly neutralizing nanobodies by phage display after immunization of dromedaries with different soluble trimeric envelope proteins derived from HIV-1 subtype C...
August 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28820969/a-disulfide-stabilized-human-vl-single-domain-antibody-library-is-a-source-of-soluble-and-highly-thermostable-binders
#20
Kevin A Henry, Hiba Kandalaft, Michael J Lowden, Martin A Rossotti, Henk van Faassen, Greg Hussack, Yves Durocher, Dae Young Kim, Jamshid Tanha
We have previously shown that incorporation of a second intradomain disulfide linkage into camelid VHH and human VH/VL single-domain antibodies confers increased thermostability. Here, we explored the effects of introducing an additional disulfide linkage, formed between Cys48 and Cys64 (Kabat numbering), into a phage-displayed synthetic human VL library. In comparison to an identical library bearing only the highly conserved Cys23-Cys88 disulfide linkage, the disulfide-stabilized VL library tolerated a similar degree of randomization but retained a higher level of functional diversity after selection with protein L...
October 2017: Molecular Immunology
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