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antibody phage display

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https://www.readbyqxmd.com/read/28193493/development-of-a-monoclonal-sandwich-elisa-for-direct-detection-of-bluetongue-virus-8-in-infected-animals
#1
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
#2
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/28186116/targeting-of-phage-particles-towards-endothelial-cells-by-antibodies-selected-through-a-multi-parameter-selection-strategy
#3
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/28184219/identification-of-bacterial-surface-antigens-by-screening-peptide-phage-libraries-using-whole-bacteria-cell-purified-antisera
#4
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/28167275/single-amino-acid-fingerprinting-of-the-human-antibody-repertoire-with-high-density-peptide-arrays
#5
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
#6
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/28154257/antibody-fragments-for-on-site-testing-of-cannabinoids-generated-via-in-vitro-affinity-maturation
#7
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/28144646/an-open-sandwich-immunoassay-for-detection-of-13-r-s-hydroxy-9-e-11-e-octadecadienoic-acid
#8
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
#9
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
#10
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
#11
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
#12
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/28109445/identification-of-novel-macropinocytosing-human-antibodies-by-phage-display-and-high-content-analysis
#13
K D Ha, S M Bidlingmaier, Y Su, N-K Lee, B Liu
Internalizing antibodies have great potential for the development of targeted therapeutics. Antibodies that internalize via the macropinocytosis pathway are particularly promising since macropinocytosis is capable of mediating rapid, bulk uptake and is selectively upregulated in many cancers. We hereby describe a method for identifying antibodies that internalize via macropinocytosis by screening phage-displayed single-chain antibody selection outputs with an automated fluorescent microscopy-based high-content analysis platform...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28107434/anti-mrka-monoclonal-antibodies-reveal-distinct-structural-and-antigenic-features-of-mrka
#14
Qun Wang, Yan Chen, Romana Cvitkovic, Meghan E Pennini, Chew Shun Chang, Mark Pelletier, Jessica Bonnell, Adem C Koksal, Herren Wu, William F Dall'Acqua, C Kendall Stover, Xiaodong Xiao
Antibody therapy against antibiotics resistant Klebsiella pneumoniae infections represents a promising strategy, the success of which depends critically on the ability to identify appropriate antibody targets. Using a target-agnostic strategy, we recently discovered MrkA as a potential antibody target and vaccine antigen. Interestingly, the anti-MrkA monoclonal antibodies isolated through phage display and hybridoma platforms all recognize an overlapping epitope, which opens up important questions including whether monoclonal antibodies targeting different MrkA epitopes can be generated and if they possess different protective profiles...
2017: PloS One
https://www.readbyqxmd.com/read/28105549/automated-antibody-de-novo-sequencing-and-its-utility-in-biopharmaceutical-discovery
#15
K Ilker Sen, Wilfred H Tang, Shruti Nayak, Yong J Kil, Marshall Bern, Berk Ozoglu, Beatrix Ueberheide, Darryl Davis, Christopher Becker
Applications of antibody de novo sequencing in the biopharmaceutical industry range from the discovery of new antibody drug candidates to identifying reagents for research and determining the primary structure of innovator products for biosimilar development. When murine, phage display, or patient-derived monoclonal antibodies against a target of interest are available, but the cDNA or the original cell line is not, de novo protein sequencing is required to humanize and recombinantly express these antibodies, followed by in vitro and in vivo testing for functional validation...
January 19, 2017: Journal of the American Society for Mass Spectrometry
https://www.readbyqxmd.com/read/28103321/derivative-of-extremophilic-50s-ribosomal-protein-l35ae-as-an-alternative-protein-scaffold
#16
Anna V Lomonosova, Andrei B Ulitin, Alexei S Kazakov, Tajib A Mirzabekov, Eugene A Permyakov, Sergei E Permyakov
Small antibody mimetics, or alternative binding proteins (ABPs), extend and complement antibody functionality with numerous applications in research, diagnostics and therapeutics. Given the superiority of ABPs, the last two decades have witnessed development of dozens of alternative protein scaffolds (APSs) for the design of ABPs. Proteins from extremophiles with their high structural stability are especially favorable for APS design. Here, a 10X mutant of the 50S ribosomal protein L35Ae from hyperthermophilic archaea Pyrococcus horikoshii has been probed as an APS...
2017: PloS One
https://www.readbyqxmd.com/read/28101463/development-of-a-novel-human-single-chain-antibody-against-egfrviii-antigen-by-phage-display-technology
#17
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Bahman Akbari, Shiva Ahdi Khosroshahi
Purpose: EGFRvIII as the most common mutant variant of the epidermal growth factor receptor is resulting from deletion of exons 2-7 in the coding sequence and junction of exons 1 and 8 through a novel glycine residue. EGFRvIII is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. The aim of the present study was identification of a novel single chain antibody against EGFRvIII as a promising target for cancer therapy. Methods: In this study, a synthetic peptide corresponding to EGFRvIII protein was used for screening a naive human scFv phage library...
December 2016: Advanced Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28095447/identification-and-characterization-of-single-chain-antibodies-that-specifically-bind-gi-noroviruses
#18
Amy M Hurwitz, Wanzhi Huang, Baijun Kou, Mary K Estes, Robert L Atmar, Timothy Palzkill
Norovirus infections commonly lead to outbreaks of acute gastroenteritis and spread quickly, resulting in many health and economic challenges prior to diagnosis. Rapid and reliable diagnostic tests are therefore essential to identify infections and to guide the appropriate clinical responses at the point-of-care. Existing tools, including RT-PCR and enzyme immunoassays, pose several limitations based on the significant time, equipment and expertise required to elicit results. Immunochromatographic assays available for use at the point-of-care have poor sensitivity and specificity, especially for genogroup I noroviruses, thus requiring confirmation of results with more sensitive testing methods...
2017: PloS One
https://www.readbyqxmd.com/read/28092036/creation-of-phosphotyrosine-superbinders-by-directed-evolution-of-an-sh2-domain
#19
Haiming Huang, Tomonori Kaneko, Sachdev S Sidhu, Shawn S C Li
Commercial antibodies raised against phosphotyrosine have been widely used as reagents to detect or isolate tyrosine-phosphorylated proteins from cellular samples. However, these antibodies are costly and are not amenable to in-house production in an academic lab setting. In this chapter, we describe a method to generate super-high affinity SH2 domains, dubbed the phosphotyrosine superbinders, by evolving a natural SH2 domain using the phage display technology. The superbinders are stable and can be easily produced in Escherichia coli in large quantities...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28090795/isolation-and-characterization-of-anti-c-met-single-chain-fragment-variable-scfv-antibodies
#20
Elmira Safaie Qamsari, Zahra Sharifzadeh, Salman Bagheri, Farhad Riazi-Rad, Vahid Younesi, Mohsen Abolhassani, Sepideh Safaei Ghaderi, Behzad Baradaran, Mohammad Hossein Somi, Mehdi Yousefi
The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques...
January 16, 2017: Journal of Immunotoxicology
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