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https://www.readbyqxmd.com/read/28211904/structural-and-functional-characterization-of-deep-sea-thermophilic-bacteriophage-gve2-hnh-endonuclease
#1
Likui Zhang, Dandan Xu, Yanchao Huang, Xinyuan Zhu, Mianwen Rui, Ting Wan, Xin Zheng, Yulong Shen, Xiangdong Chen, Kesen Ma, Yong Gong
HNH endonucleases in bacteriophages play a variety of roles in the phage lifecycle as key components of phage DNA packaging machines. The deep-sea thermophilic bacteriophage Geobacillus virus E2 (GVE2) encodes an HNH endonuclease (GVE2 HNHE). Here, the crystal structure of GVE2 HNHE is reported. This is the first structural study of a thermostable HNH endonuclease from a thermophilic bacteriophage. Structural comparison reveals that GVE2 HNHE possesses a typical ββα-metal fold and Zn-finger motif similar to those of HNH endonucleases from other bacteriophages, apart from containing an extra α-helix, suggesting conservation of these enzymes among bacteriophages...
February 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28208072/identification-of-triosephosphate-isomerase-as-a-novel-allergen-in-octopus-fangsiao
#2
Yang Yang, Zhong-Wei Chen, Barry K Hurlburt, Gui-Ling Li, Yong-Xia Zhang, Dan-Xia Fei, Hai-Wang Shen, Min-Jie Cao, Guang-Ming Liu
Octopus is an important mollusk in human dietary for its nutritional value, however it also causes allergic reactions in humans. Major allergens from octopus have been identified, while the knowledge of novel allergens remains poor. In the present study, a novel allergen with molecular weight of 28kDa protein was purified from octopus (Octopus fangsiao) and identified as triosephosphate isomerase (TIM) by mass spectrometry. TIM aggregated beyond 45°C, and its IgE-binding activity was affected under extreme pH conditions due to the altered secondary structure...
February 13, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28193493/development-of-a-monoclonal-sandwich-elisa-for-direct-detection-of-bluetongue-virus-8-in-infected-animals
#3
Andre Ten Haaf, Johannes Kohl, Sibylle Pscherer, Hans-Peter Hamann, Hans Ulrich Eskens, Max Bastian, Stefan Gattenlöhner, Mehmet Kemal Tur
Bluetongue is an infectious viral disease which can cause mortality in affected ruminants, and tremendous economic damage via impacts upon fertility, milk production and the quality of wool. The disease is caused by bluetongue virus (BTV) which is transmitted by species of Culicoides biting midge. Rapid detection of BTV is required to contain disease outbreaks and reduce economic losses. The purpose of this study was to develop a monoclonal sandwich ELISA for direct detection of BTV in infected animals. Phage display technology was used to isolate BTV specific antibody fragments by applying the human scFv Tomlinson antibody libraries directly on purified BTV-8 particles...
February 10, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28186432/development-of-anti-cd47-single-chain-variable-fragment-targeted-magnetic-nanoparticles-for-treatment-of-human-bladder-cancer
#4
Gashin Rezaei, Mahdi Habibi-Anbouhi, Morteza Mahmoudi, Kayhan Azadmanesh, Shima Moradi-Kalbolandi, Mahdi Behdani, Leila Ghazizadeh, Mohsen Abolhassani, Mohammad Ali Shokrgozar
AIM: To develop a novel anti-CD47 single-chain variable fragment (scFv) functionalized magnetic nanoparticles (MNPs) for targeting bladder cell lines and its applicability in thermotherapy. MATERIAL & METHODS: An immunized murine antibody phage display library was constructed and screened to isolate anti-CD47 binders. A scFv was selected and conjugated to MNPs which was then utilized to discriminate CD47(+) bladder cells along with assessing its efficacy in thermotherapy...
February 10, 2017: Nanomedicine
https://www.readbyqxmd.com/read/28186164/a-new-approach-to-quantification-of-mab-aggregates-using-peptide-affinity-probes
#5
Crystal S F Cheung, Kyle W Anderson, Pooja M Patel, Keale L Cade, Karen W Phinney, Illarion V Turko
Using mAbs as therapeutic molecules is complicated by the propensity of mAbs to aggregate at elevated concentrations, which can lead to a variety of adverse events in treatment. Here, we describe a proof-of-concept for new methodology to detect and quantify mAb aggregation. Assay development included using an aggregated mAb as bait for screening of phage display peptide library and identifying those peptides with random sequence which can recognize mAb aggregates. Once identified, the selected peptides can be used for developing quantitative methods to assess mAb aggregation...
February 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28186116/targeting-of-phage-particles-towards-endothelial-cells-by-antibodies-selected-through-a-multi-parameter-selection-strategy
#6
Ole A Mandrup, Simon Lykkemark, Peter Kristensen
One of the hallmarks of cancer is sustained angiogenesis. Here, normal endothelial cells are activated, and their formation of new blood vessels leads to continued tumour growth. An improved patient condition is often observed when angiogenesis is prevented or normalized through targeting of these genomically stable endothelial cells. However, intracellular targets constitute a challenge in therapy, as the agents modulating these targets have to be delivered and internalized specifically to the endothelial cells...
February 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28185387/engineering-sortase-a-by-screening-a-second-generation-library-using-phage-display
#7
Lena Schmohl, Jan Bierlmeier, Fabian Gerth, Christian Freund, Dirk Schwarzer
Sortase-mediated ligation is one of the most commonly used chemo-enzymatic techniques for the site-specific modification of proteins. We have established a new library of sortase mutants for directed evolution of sortase substrate selectivity. Phage display screens of this second-generation library yielded sortase mutants that ligate substrate proteins containing an APxTG or FPxTG recognition sequence instead of the canonical LPxTG sorting motif. These findings indicate that the second-generation sortase library is well suited for sortase engineering in order to increase the versatility of sortase-mediated ligation...
February 10, 2017: Journal of Peptide Science: An Official Publication of the European Peptide Society
https://www.readbyqxmd.com/read/28184219/identification-of-bacterial-surface-antigens-by-screening-peptide-phage-libraries-using-whole-bacteria-cell-purified-antisera
#8
Yun-Fei Hu, Dun Zhao, Xing-Long Yu, Yu-Li Hu, Run-Cheng Li, Meng Ge, Tian-Qi Xu, Xiao-Bo Liu, Hua-Yuan Liao
Bacterial surface proteins can be good vaccine candidates. In the present study, we used polyclonal antibodies purified with intact Erysipelothrix rhusiopthiae to screen phage-displayed random dodecapeptide and loop-constrained heptapeptide libraries, which led to the identification of mimotopes. Homology search of the mimotope sequences against E. rhusiopthiae-encoded ORF sequences revealed 14 new antigens that may localize on the surface of E. rhusiopthiae. When these putative surface proteins were used to immunize mice, 9/11 antigens induced protective immunity...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28167526/vibrio-phage-kvp40-encodes-a-functional-nad-salvage-pathway
#9
Jae Yun Lee, Zhiqun Li, Eric S Miller
: The genome of T4-type Vibrio bacteriophage KVP40 has five genes predicted to encode proteins of pyridine nucleotide metabolism, of which two, nadV and natV, would suffice for an NAD(+) salvage pathway. NadV is an apparent nicotinamide phosphoribosyltransferase (NAmPRTase) and NatV is an apparent bifunctional nicotinamide mononucleotide adenylyltransferase (NMNATase) and nicotinamide-adenine dinucleotide pyrophosphatase (Nudix hydrolase). Genes encoding the predicted salvage pathway were cloned, expressed in E...
February 6, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28167275/single-amino-acid-fingerprinting-of-the-human-antibody-repertoire-with-high-density-peptide-arrays
#10
Laura K Weber, Andrea Palermo, Jonas Kügler, Olivier Armant, Awale Isse, Simone Rentschler, Thomas Jaenisch, Jürgen Hubbuch, Stefan Dübel, Alexander Nesterov-Mueller, Frank Breitling, Felix F Loeffler
The antibody species that patrol in a patient's blood are an invaluable part of the immune system. While most of them shield us from life-threatening infections, some of them do harm in autoimmune diseases. If we knew exactly all the antigens that elicited all the antibody species within a group of patients, we could learn which ones correlate with immune protection, are irrelevant, or do harm. Here, we demonstrate an approach to this question: First, we use a plethora of phage-displayed peptides to identify many different serum antibody binding peptides...
February 3, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28165018/recombinant-phage-elicits-protective-immune-response-against-systemic-s-globosa-infection-in-mouse-model
#11
Feng Chen, Rihua Jiang, Yicun Wang, Mingji Zhu, Xu Zhang, Shuai Dong, Hongxi Shi, Li Wang
Sporothrix globosa is a type of fungus that typically infects immunocompromised patients. Its prevention continues to pose a challenge. A 70-KDa glycoprotein (Gp70) of Sporothrix has been previously reported to protect host against infection from this fungus. Here, we displayed an epitope peptide (kpvqhalltplgldr) of Gp70 on the major coat protein (pIII), and investigated its efficiency as a vaccine for preventing S. globosa infection. The recombinant phage and the heat-killed S. globosa were used to immunize mice separately...
February 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28165000/high-affinity-anchoring-of-the-decoration-protein-pb10-onto-the-bacteriophage-t5-capsid
#12
Emeline Vernhes, Madalena Renouard, Bernard Gilquin, Philippe Cuniasse, Dominique Durand, Patrick England, Sylviane Hoos, Alexis Huet, James F Conway, Anatoly Glukhov, Vladimir Ksenzenko, Eric Jacquet, Naïma Nhiri, Sophie Zinn-Justin, Pascale Boulanger
Bacteriophage capsids constitute icosahedral shells of exceptional stability that protect the viral genome. Many capsids display on their surface decoration proteins whose structure and function remain largely unknown. The decoration protein pb10 of phage T5 binds at the centre of the 120 hexamers formed by the major capsid protein. Here we determined the 3D structure of pb10 and investigated its capsid-binding properties using NMR, SAXS, cryoEM and SPR. Pb10 consists of an α-helical capsid-binding domain and an Ig-like domain exposed to the solvent...
February 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28154576/regulatory-shifts-in-plastid-transcription-play-a-key-role-in-morphological-conversions-of-plastids-during-plant-development
#13
REVIEW
Monique Liebers, Björn Grübler, Fabien Chevalier, Silva Lerbs-Mache, Livia Merendino, Robert Blanvillain, Thomas Pfannschmidt
Plastids display a high morphological and functional diversity. Starting from an undifferentiated small proplastid, these plant cell organelles can develop into four major forms: etioplasts in the dark, chloroplasts in green tissues, chromoplasts in colored flowers and fruits and amyloplasts in roots. The various forms are interconvertible into each other depending on tissue context and respective environmental condition. Research of the last two decades uncovered that each plastid type contains its own specific proteome that can be highly different from that of the other types...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28154257/antibody-fragments-for-on-site-testing-of-cannabinoids-generated-via-in-vitro-affinity-maturation
#14
Izumi Morita, Hiroyuki Oyama, Mayumi Yasuo, Kazuhisa Matsuda, Kengo Katagi, Aya Ito, Hiroka Tatsuda, Hiroyuki Tanaka, Satoshi Morimoto, Norihiro Kobayashi
Law enforcement against illicit use of cannabis and related substances requires rapid, feasible, and reliable tools for on-site testing of cannabinoids. Notably, methods based on cannabinoid-specific antibodies enable efficient screening of multiple specimens. Antibody engineering may accelerate development of modern and robust testing systems. Here, we used in vitro affinity maturation to generate a single-chain Fv fragment (scFv) that recognizes with high affinity the psychoactive cannabinoid, Δ(9)-tetrahydrocannabinol (THC)...
2017: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28153726/k-ras-g12d-selective-inhibitory-peptides-generated-by-random-peptide-t7-phage-display-technology
#15
Kotaro Sakamoto, Yusuke Kamada, Tomoya Sameshima, Masahiro Yaguchi, Ayumu Niida, Shigekazu Sasaki, Masanori Miwa, Shoichi Ohkubo, Jun-Ichi Sakamoto, Masahiro Kamaura, Nobuo Cho, Akiyoshi Tani
Amino-acid mutations of Gly(12) (e.g. G12D, G12V, G12C) of V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-Ras), the most promising drug target in cancer therapy, are major growth drivers in various cancers. Although over 30 years have passed since the discovery of these mutations in most cancer patients, effective mutated K-Ras inhibitors have not been marketed. Here, we report novel and selective inhibitory peptides to K-Ras(G12D). We screened random peptide libraries displayed on T7 phage against purified recombinant K-Ras(G12D), with thorough subtraction of phages bound to wild-type K-Ras, and obtained KRpep-2 (Ac-RRCPLYISYDPVCRR-NH2) as a consensus sequence...
January 30, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28144646/an-open-sandwich-immunoassay-for-detection-of-13-r-s-hydroxy-9-e-11-e-octadecadienoic-acid
#16
Jinhua Dong, Mototada Shichiri, Chan-I Chung, Takahiro Shibata, Koji Uchida, Yoshihisa Hagihara, Yasukazu Yoshida, Hiroshi Ueda
Lipid peroxidation is involved in many disorders and diseases such as cardiovascular disease, cancers, neurodegenerative diseases, and even aging. Lipid peroxidation products existing in blood or bodily fluids are very important biomarkers for the diagnosis of such diseases. In particular, 13(R,S)-hydroxy-9(E),11(E)-octadecadienoic acid (13-(E,E)-HODE) is an oxidiation product of linoleic acid, which is an important biomarker for many diseases such as diabetes and Alzheimer's disease. In this study, we successfully displayed the antigen-binding fragment of an antibody produced by hybridoma 1213-1 on the M13 phage and performed analysis of the antibody variable region genes...
February 1, 2017: Analyst
https://www.readbyqxmd.com/read/28128766/isolation-and-characterization-of-anti-ror1-single-chain-fragment-variable-antibodies-using-phage-display-technique
#17
Leili Aghebati Maleki, Vahid Younesi, Farhad Jadidi-Niaragh, Behzad Baradaran, Jafar Majidi, Mehdi Yousefi
Receptor tyrosine kinase-like orphan receptor (ROR1) belongs to one of the families of receptor tyrosine kinases (RTKs). RTKs are involved in the various physiologic cellular functions including proliferation, migration, survival, signaling and differentiation. Several RTKs are deregulated in various cancers implying the targeting potential of these molecules in cancer therapy. ROR1 has recently been shown to be expressed in various types of cancer cells but not in normal adult cells. Hence a molecular inhibitor of extracellular domain of ROR1 that inhibits ROR1-cell surface interaction is of great therapeutic importance...
January 20, 2017: Human Antibodies
https://www.readbyqxmd.com/read/28125612/a-recombinant-human-anti-platelet-scfv-antibody-produced-in-pichia-pastoris-for-atheroma-targeting
#18
Amelie Vallet-Courbin, Mélusine Larivière, Agnès Hocquellet, Audrey Hemadou, Sarjapura-Nagaraja Parimala, Jeanny Laroche-Traineau, Xavier Santarelli, Gisèle Clofent-Sanchez, Marie-Josée Jacobin-Valat, Abdelmajid Noubhani
Cells of the innate and adaptive immune system are key factors in the progression of atherosclerotic plaque, leading to plaque instability and rupture, potentially resulting in acute atherothrombotic events such as coronary artery disease, cerebrovascular disease and peripheral arterial disease. Here, we describe the cloning, expression, purification, and immunoreactivity assessment of a recombinant single-chain variable fragment (scFv) derived from a human anti-αIIbβ3 antibody (HuAb) selected to target atheromatous lesions for the presence of platelets...
2017: PloS One
https://www.readbyqxmd.com/read/28122516/tdp-43-protein-variants-as-biomarkers-in-amyotrophic-lateral-sclerosis
#19
Stephanie M Williams, Galam Khan, Brent T Harris, John Ravits, Michael R Sierks
BACKGROUND: TDP-43 aggregates accumulate in individuals affected by amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases, representing potential diagnostic and therapeutic targets. Using an atomic force microscopy based biopanning protocol developed in our lab, we previously isolated 23 TDP-43 reactive antibody fragments with preference for human ALS brain tissue relative to frontotemporal dementia, a related neurodegeneration, and healthy samples from phage-displayed single chain antibody fragment (scFv) libraries...
January 25, 2017: BMC Neuroscience
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
#20
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
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