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Grace VanHoy, Michelle Carman, Greg Habing, Jeffrey Lakritz, C Austin Hinds, Andrew Niehaus, Ray M Kaplan, Antoinette E Marsh
Haemonchosis in camelids remains a challenging disease to treat, and prevention has become increasingly problematic due to widespread anthelmintic resistance. Barbervax® is an adjuvanted vaccine containing natural H-11, H-gal-GP antigens obtained from Haemonchus contortus adults via a proprietary process and solubilized in Quil A. This vaccine is approved for use in Australia, after demonstrating its safety and efficacy in sheep and goats. There are no published studies evaluating Barbervax in other ruminants/pseudoruminants such as camelids which can be parasitized with H...
March 15, 2018: Veterinary Parasitology
Fatemeh Faraji, Nader Tajik, Mahdi Behdani, Mohammad Ali Shokrgozar, Amir Hassan Zarnani, Fatemeh Shahhosseini, Mahdi Habibi-Anbouhi
CD22 is a B-cell-specific trans-membrane glycoprotein which is found on the surface of the most B-cells and modulates their function, survival, and apoptosis. Recently, targeting this cell surface biomarker in B-cell malignancies and disorders has attracted a lot of attention. The variable domain of camelid single chain antibodies (VHH, Nanobody) is a form of antibodies with novel properties including small size (15-17 kDa), thermal and chemical stability, high affinity and homology to human antibody sequences...
March 15, 2018: Biotechnology and Applied Biochemistry
Almut Prkno, Matthias Kaiser, Daniela Goerigk, Martin Pfeffer, Thomas W Vahlenkamp, Donata Hoffmann, Martin Beer, Alexander Starke
Cowpox virus (CPXV) infection is a reportable and potentially zoonotic disease that occurs sporadically in a variety of animals. During the past six decades, CPXV infection has been extensively researched and described in both domestic (cat, dog, horse, cattle) and zoo animals (e. g. elephant, rhinoceros, okapi). Of note, a review of the literature produced only three reports of CPXV in individual or small groups of South American camelids. The goal of this review was to describe the current knowledge as it relates to clinical features of CPXV infection in South American camelids and to compare the clinical manifestations with those described in other animal species...
February 2018: Tierärztliche Praxis. Ausgabe G, Grosstiere/Nutztiere
Andreas H Laustsen, José María Gutiérrez, Cecilie Knudsen, Kristoffer H Johansen, Erick Bermúdez Méndez, Felipe A Cerni, Jonas A Jürgensen, Mia Øhlenschlæger, Line Ledsgaard, Andrea Martos Esteban, Urska Pus, Mikael R Andersen, Bruno Lomonte, Mikael Engmark, Manuela B Pucca
Antibody technologies are being increasingly applied in the field of toxinology. Fuelled by the many advances in immunology, synthetic biology, and antibody research, different approaches and antibody formats are being investigated for the ability to neutralize animal toxins. These different molecular formats each have their own therapeutic characteristics. In this review, we provide an overview of the advances made in the development of toxin-targeting antibodies, and discuss the benefits and drawbacks of different antibody formats in relation to their ability to neutralize toxins, pharmacokinetic features, propensity to cause adverse reactions, formulation, and expression for research and development (R&D) purposes and large-scale manufacturing...
March 10, 2018: Toxicon: Official Journal of the International Society on Toxinology
Fatemeh Kazemi-Lomedasht, Serge Muyldermans, Mahdi Habibi-Anbouhi, Mahdi Behdani
Objectives: Nanobodies, the single domain antigen binding fragments of heavy chain-only antibodies occurring naturally in camelid sera, are the smallest intact antigen binding entities. Their minimal size assists in reaching otherwise largely inaccessible regions of antigens. However, their camelid origin raises a possible concern of immunogenicity when used for human therapy. Humanization is a promising approach to overcome the problem. Materials and Methods: Here, we designed a humanized version of previously developed nanobody (anti vascular endothelial growth factor nanobody), evaluated and compared its predicted 3D structure, affinity and biological activity with its original wild type nanobody...
March 2018: Iranian Journal of Basic Medical Sciences
Muhammad A Saeed, Mohammed H Rashid, Jane Vaughan, Abdul Jabbar
Members of the genus Sarcocystis (Apicomplexa: Sarcocystidae) are intracellular protozoan parasites that infect a wide range of domestic and wild animals, resulting in economic losses in production animals worldwide. Sarcocystis spp. have indirect life-cycles where canids and felids serve as main definitive hosts while a range of domestic and wild animals serve as intermediate hosts, including South American camelids (SACs) such as alpacas, llamas and guanacos. These animals primarily occur in South American countries on Andean, elevated plains but in recent years, alpacas and llamas have become emerging animal industries in other parts of the world such as Australia, Europe and the USA due to their high-quality fiber, meat and hides...
March 6, 2018: Parasites & Vectors
Robert Alvin Bernedo-Navarro, Ema Romão, Tomomasa Yano, Joar Pinto, Henri De Greve, Yann G-J Sterckx, Serge Muyldermans
BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) are a subset of pathogens leading to illnesses such as diarrhea, hemolytic uremic syndrome and even death. The Shiga toxins are the main virulence factors and divided in two groups: Stx1 and Stx2, of which the latter is more frequently associated with severe pathologies in humans. RESULTS: An immune library of nanobodies (Nbs) was constructed after immunizing an alpaca with recombinant Shiga toxin-2a B subunit (rStx2aB), to retrieve multiple rStx2aB-specific Nbs...
March 1, 2018: Toxins
Pamela Holzlöhner, Monique Butze, Natalia Maier, Nicole Hebel, Erik Schliebs, Burkhard Micheel, Jonas Füner, Gabriele Heidicke, Katja Hanack
Camelids possess antibodies with a conventional four-chain structure consisting of two heavy and two light chains (of subclass IgG1) but further they also generate heavy-chain only antibodies (of subclass IgG2 and 3) which are fully functional in antigen binding. In this study subclass-specific murine monoclonal antibodies specific to conventional camelid IgG1 and heavy-chain only IgG2/3 were generated and validated for the use as potent secondary detection reagents. The monoclonal antibodies are able to differentiate between all camelid IgGs, conventional four-chain camelid antibodies (of subclass IgG1) and exclusively heavy chain-only antibodies (of subclasses IgG2 and IgG3)...
March 2018: Veterinary Immunology and Immunopathology
Marylène Vandevenne, Mathieu Dondelinger, Sami Yunus, Astrid Freischels, Régine Freischels, Oscar Crasson, Noureddine Rhazi, Pierre Bogaerts, Moreno Galleni, Patrice Filée
Biosensors are becoming increasingly important and implemented in various fields such as pathogen detection, molecular diagnosis, environmental monitoring, and food safety control. In this context, we used β-lactamases as efficient reporter enzymes in several protein-protein interaction studies. Furthermore, their ability to accept insertions of peptides or structured proteins/domains strongly encourages the use of these enzymes to generate chimeric proteins. In a recent study, we inserted a single-domain antibody fragment into the Bacillus licheniformis BlaP β-lactamase...
February 1, 2018: Journal of Visualized Experiments: JoVE
Masahito Yamagata, Joshua R Sanes
Sensitive and specific antibodies are essential for detecting molecules in cells and tissues. However, currently used polyclonal and monoclonal antibodies are often less specific than desired, difficult to produce, and available in limited quantities. A promising recent approach to circumvent these limitations is to employ chemically defined antigen-combining domains called "nanobodies," derived from single-chain camelid antibodies. Here, we used nanobodies to prepare sensitive unimolecular detection reagents by genetically fusing cDNAs encoding nanobodies to enzymatic or antigenic reporters...
February 13, 2018: Proceedings of the National Academy of Sciences of the United States of America
Sunanda Singh, Genoveva Murillo, Dong Chen, Ashutosh S Parihar, Rajendra G Mehta
Background: The serendipitous discovery of heavy-chain antibodies devoid of light chains in camelids and the subsequent development of VHHs (variable region of camelid heavy chain) have provided a very important tool for research and possibly for therapeutics. In this study, we synthesized single-domain 15-kDa antibody SBT-100 (anti-STAT3 B VHH13) against human STAT3 (signal transducer and activator of transcription) that binds selectively to STAT3 and suppresses the function of phosphorylated STAT3 (p-STAT3)...
2018: Breast Cancer: Basic and Clinical Research
Conor McMahon, Alexander S Baier, Roberta Pascolutti, Marcin Wegrecki, Sanduo Zheng, Janice X Ong, Sarah C Erlandson, Daniel Hilger, Søren G F Rasmussen, Aaron M Ring, Aashish Manglik, Andrew C Kruse
Camelid single-domain antibody fragments ('nanobodies') provide the remarkable specificity of antibodies within a single 15-kDa immunoglobulin VHH domain. This unique feature has enabled applications ranging from use as biochemical tools to therapeutic agents. Nanobodies have emerged as especially useful tools in protein structural biology, facilitating studies of conformationally dynamic proteins such as G-protein-coupled receptors (GPCRs). Nearly all nanobodies available to date have been obtained by animal immunization, a bottleneck restricting many applications of this technology...
March 2018: Nature Structural & Molecular Biology
B Schauer, R Krametter-Frötscher, F Knauer, R Ehricht, S Monecke, A T Feßler, S Schwarz, T Grunert, J Spergser, I Loncaric
The aim of this study was to determine the prevalence, the antimicrobial resistance patterns and the genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) from Austrian ruminants and New World camelids that were treated at the University of Veterinary Medicine, Vienna. Between April 2014 and January 2017, 723 nasal swabs originating from ruminants and New World camelids were examined. MRSA isolates were characterized by mecA/mecA1/mecC PCRs and by DNA microarray analysis. They were genotyped by spa typing, dru typing, MLST and MLVA...
February 2018: Veterinary Microbiology
Amanda Sparkes, Patrick De Baetselier, Lea Brys, Inês Cabrito, Yann G-J Sterckx, Steve Schoonooghe, Serge Muyldermans, Geert Raes, Richard Bucala, Peter Vanlandschoot, Jo A Van Ginderachter, Benoît Stijlemans
Sepsis-leading to septic shock-is the leading cause of death in intensive care units. The systemic inflammatory response to infection, which is initiated by activated myeloid cells, plays a key role in the lethal outcome. Macrophage migration inhibitory factor (MIF) is an upstream immunoregulatory mediator, released by myeloid cells, that underlies a common genetic susceptibility to different infections and septic shock. Accordingly, strategies that are aimed at inhibiting the action of MIF have therapeutic potential...
January 25, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Naoya Shinozaki, Ryuji Hashimoto, Masanori Noda, Susumu Uchiyama
Recently, we showed that immunized rabbit heavy chain variable regions (rVHs) can have strong antigen binding activity comparable to that of the camelid variable domain of the heavy chain of heavy chain antibody (VHH). These rVHs lack the light chain variable regions (rVLs), which exist in the authentic Fab format; thus, molecular surfaces at the interface region of rVHs are exposed to solvent. This physical feature may change physicochemical properties, such as causing reduced stability. By overcoming potential physicochemical issues through engineering the interface region, rVHs could become more useful as single-domain antibodies...
February 1, 2018: Journal of Bioscience and Bioengineering
Traian Sulea, Greg Hussack, Shannon Ryan, Jamshid Tanha, Enrico O Purisima
Assisted Design of Antibody and Protein Therapeutics (ADAPT) is an affinity maturation platform interleaving predictions and testing that was previously validated on monoclonal antibodies (mAbs). This study expands the applicability of ADAPT to single-domain antibodies (sdAbs), a promising class of recombinant antibody-based biologics. As a test case, we used the camelid sdAb A26.8, a VHH that binds Clostridium difficile toxin A (TcdA) relatively weakly but displays a reasonable level of TcdA neutralization...
February 2, 2018: Scientific Reports
Polina L Perelman, Rudolf Pichler, Anna Gaggl, Denis M Larkin, Terje Raudsepp, Fahad Alshanbari, Heather M Holl, Samantha A Brooks, Pamela A Burger, Kathiravan Periasamy
The availability of genomic resources including linkage information for camelids has been very limited. Here, we describe the construction of a set of two radiation hybrid (RH) panels (5000RAD and 15000RAD) for the dromedary (Camelus dromedarius) as a permanent genetic resource for camel genome researchers worldwide. For the 5000RAD panel, a total of 245 female camel-hamster radiation hybrid clones were collected, of which 186 were screened with 44 custom designed marker loci distributed throughout camel genome...
January 31, 2018: Scientific Reports
Abdullah Hoter, Mahdi Amiri, Abdelbary Prince, Hassan Amer, Mohamad Warda, Hassan Y Naim
Increased expression of heat shock proteins (HSPs) following heat stress or other stress conditions is a common physiological response in almost all living organisms. Modification of cytosolic proteins including HSPs by O-GlcNAc has been shown to enhance their capabilities for counteracting lethal levels of cellular stress. Since HSPs are key players in stress resistance and protein homeostasis, we aimed to analyze their forms at the cellular and molecular level using camel and human HSPs as models for efficient and moderate thermotolerant mammals, respectively...
January 30, 2018: International Journal of Molecular Sciences
Ryota Saito, Yutaro Saito, Hikaru Nakazawa, Takamitsu Hattori, Izumi Kumagai, Mitsuo Umetsu, Koki Makabe
Biomolecules which recognize inorganic materials and metal surfaces gain much attention for creating new type of nanomaterials and sensors. 4F2, a camelid VHH antibody, recognizes ZnO surface and has been applied for sensor applications. 4F2 was constructed sequential CDR replacement on the parental VHH antibody, termed the Construction of Antibody by Integrating Grafting and Evolution Technology; CAnIGET procedure. Here, we evaluate the influence of CDR replacements during 4F2 generation using calorimetric technique...
January 22, 2018: Journal of Biochemistry
Antony S K Yerabham, Andreas Müller-Schiffmann, Tamar Ziehm, Andreas Stadler, Sabrina Köber, Xela Indurkhya, Rita Marreiros, Svenja V Trossbach, Nicholas J Bradshaw, Ingrid Prikulis, Dieter Willbold, Oliver H Weiergräber, Carsten Korth
Accumulating evidence suggests an important role for the Disrupted-in-Schizophrenia 1 (DISC1) protein in neurodevelopment and chronic mental illness. In particular, the C-terminal 300 amino acids of DISC1 have been found to mediate important protein-protein interactions and to harbor functionally important phosphorylation sites and disease-associated polymorphisms. However, long disordered regions and oligomer-forming subdomains have so far impeded structural analysis. VHH domains derived from camelid heavy chain only antibodies are minimal antigen binding modules with appreciable solubility and stability, which makes them well suited for the stabilizing proteins prior to structural investigation...
2018: PloS One
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