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https://www.readbyqxmd.com/read/28218671/human-scfvs-that-counteract-bioactivities-of-staphylococcus-aureus-tsst-1
#1
Thunchanok Rukkawattanakul, Nitat Sookrung, Watee Seesuay, Nattawat Onlamoon, Pornphan Diraphat, Wanpen Chaicumpa, Nitaya Indrawattana
Some Staphylococcus aureus isolates produced toxic shock syndrome toxin-1 (TSST-1) which is a pyrogenic toxin superantigen (PTSAg). The toxin activates a large fraction of peripheral blood T lymphocytes causing the cells to proliferate and release massive amounts of pro-inflammatory cytokines leading to a life-threatening multisystem disorder: toxic shock syndrome (TSS). PTSAg-mediated-T cell stimulation circumvents the conventional antigenic peptide presentation to T cell receptor (TCR) by the antigen-presenting cell (APC)...
February 17, 2017: Toxins
https://www.readbyqxmd.com/read/28202953/fully-human-cd19-specific-chimeric-antigen-receptors-for-t-cell-therapy
#2
D Sommermeyer, T Hill, S M Shamah, A I Salter, Y Chen, K M Mohler, S R Riddell
Impressive results have been achieved by adoptively transferring T-cells expressing CD19-specific CARs with binding domains from murine mAbs to treat B-cell malignancies. T-cell mediated immune responses specific for peptides from the murine scFv antigen-binding domain of the CAR can develop in patients and result in premature elimination of CAR-T-cells increasing the risk of tumor relapse. As fully human scFv might reduce immunogenicity, we generated CD19-specific human scFvs with similar binding characteristics as the murine FMC63-derived scFv using human Ab/DNA-libraries...
February 16, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/28193211/display-of-single-chain-variable-fragments-on-bacteriophage-ms2-virus-like-particles
#3
Christopher A Lino, Jerri C Caldeira, David S Peabody
BACKGROUND: Virus-like particles (VLPs) of the RNA bacteriophage MS2 have many potential applications in biotechnology. MS2 VLPs provide a platform for peptide display and affinity selection (i.e. biopanning). They are also under investigation as vehicles for targeted drug delivery, using display of receptor-specific peptides or nucleic acid aptamers to direct their binding to specific cell-surface receptors. However, there are few molecules more suited to the precise targeting and binding of a cellular receptor than antibodies...
February 13, 2017: Journal of Nanobiotechnology
https://www.readbyqxmd.com/read/28165915/humanization-of-rabbit-monoclonal-antibodies-via-grafting-combined-kabat-imgt-paratome%C3%A2-complementarity-determining-regions-rationale-and-examples
#4
Yi-Fan Zhang, Mitchell Ho
Rabbit monoclonal antibodies (RabMAbs) can recognize diverse epitopes, including those poorly immunogenic in mice and humans. However, there have been only a few reports on RabMAb humanization, an important antibody engineering step usually done before clinical applications are investigated. To pursue a general method for humanization of RabMAbs, we analyzed the complex structures of five RabMAbs with their antigens currently available in the Protein Data Bank, and identified antigen-contacting residues on the rabbit Fv within the 6 Angstrom distance to its antigen...
February 6, 2017: MAbs
https://www.readbyqxmd.com/read/28128766/isolation-and-characterization-of-anti-ror1-single-chain-fragment-variable-antibodies-using-phage-display-technique
#5
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/28122516/tdp-43-protein-variants-as-biomarkers-in-amyotrophic-lateral-sclerosis
#6
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/28095447/identification-and-characterization-of-single-chain-antibodies-that-specifically-bind-gi-noroviruses
#7
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/28075428/balanced-secretion-of-anti-cea-x-anti-cd3-diabody-chains-using-the-2a-self-cleaving-peptide-maximizes-diabody-assembly-and-tumor-specific-cytotoxicity
#8
K Mølgaard, M Compte, N Nuñez-Prado, S L Harwood, L Sanz, L Alvarez-Vallina
Adoptive transfer of genetically engineered human cells secreting bispecific T cell engagers has shown encouraging therapeutic effects in preclinical models of cancer. However, reducing the toxicity and improving the effectiveness of this emerging immunotherapeutic strategy will be critical to its successful application. We have demonstrated that for gene-based bispecific antibody strategies, two-chain diabodies have a better safety profile than single-chain tandem scFvs, because their reduced tendency to form aggregates reduces the risk of inducing antigen-independent T cell activation...
January 11, 2017: Gene Therapy
https://www.readbyqxmd.com/read/28059445/computational-de-novo-design-of-antibodies-binding-to-a-peptide-with-high-affinity
#9
Venkata Giridhar Poosarla, Tong Li, Boon Chong Goh, Klaus Schulten, Thomas K Wood, Costas D Maranas
Antibody drugs play a critical role in infectious diseases, cancer, autoimmune diseases, and inflammation. However, experimental methods for the generation of therapeutic antibodies such as using immunized mice or directed evolution remain time consuming and cannot target a specific antigen epitope. Here, we describe the application of a computational framework called OptMAVEn combined with molecular dynamics to de novo design antibodies. Our reference system is antibody 2D10, a single-chain antibody (scFv) that recognizes the dodecapeptide DVFYPYPYASGS, a peptide mimic of mannose-containing carbohydrates...
January 6, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28055297/a-strategy-to-identify-linker-based-modules-for-the-allosteric-regulation-of-antibody-antigen-binding-affinities-of-different-scfvs
#10
Sarah-Jane Kellmann, Stefan Dübel, Holger Thie
Antibody single-chain variable fragments (scFvs) are used in a variety of applications, such as for research, diagnosis and therapy. Essential for these applications is the extraordinary specificity, selectivity and affinity of antibody paratopes, which can also be employed for efficient protein purification. However, this use is hampered by the high affinity for the protein to be purified because harsh elution conditions, which may impair folding, integrity or viability of the eluted biomaterials, are typically required...
January 5, 2017: MAbs
https://www.readbyqxmd.com/read/27984091/the-bacillus-subtilis-tatadcd-system-exhibits-an-extreme-level-of-substrate-selectivity
#11
Kelly M Frain, Alexander S Jones, Ronald Schoner, Kelly L Walker, Colin Robinson
The Tat system preferentially transports correctly folded proteins across the bacterial membrane although little is known of the proofreading mechanism. Most research has focused on TatABC systems from Gram-negative bacteria, especially Escherichia coli, and much less is known of the TatAC-type systems from Gram-positive organisms. We have previously shown that the Bacillus subtilis TatAdCd system is functional in an E. coli tat null background and able to transport TorA-GFP and native TorA (TMAO reductase); here, we examined its ability to transport other proteins bearing a TorA signal sequence...
October 29, 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27888111/neutralizing-human-recombinant-antibodies-against-herpes-simplex-virus-type-1-glycoproteins-b-from-a-phage-displayed-scfv-antibody-library
#12
Vahid Bagheri, Foroogh Nejatollahi, Seyed Alireza Esmaeili, Amir Abbas Momtazi, Mohamad Motamedifar, Amirhossein Sahebkar
The HSV-1 envelope glycoprotein B (gB) plays a critical role in virus entry into host cells. Neutralizing antibodies can therefore potentially prevent virus entry into target cells and cell-to-cell spread of infection. Our present study focused on the selection of neutralizing single-chain Fv (scFv) antibodies of a phage-displayed nonimmune human scFv antibody library against gB of HSV-1. To enrich specific scFvs, two phage antibodies were isolated against amino acid residues 31-43 derived from the N-terminal part of gB using panning technique...
January 15, 2017: Life Sciences
https://www.readbyqxmd.com/read/27879646/integrating-scfv-into-xmap-assays-for-the-detection-of-marine-toxins
#13
Lisa C Shriver-Lake, Jinny L Liu, P Audrey Brozozog Lee, Ellen R Goldman, Richard Dietrich, Erwin Märtlbauer, George P Anderson
Marine toxins, such as saxitoxin and domoic acid are associated with algae blooms and can bioaccumulate in shell fish which present both health and economic concerns. The ability to detect the presence of toxin is paramount for the administration of the correct supportive care in case of intoxication; environmental monitoring to detect the presence of toxin is also important for prevention of intoxication. Immunoassays are one tool that has successfully been applied to the detection of marine toxins. Herein, we had the variable regions of two saxitoxin binding monoclonal antibodies sequenced and used the information to produce recombinant constructs that consist of linked heavy and light variable domains that make up the binding domains of the antibodies (scFv)...
November 21, 2016: Toxins
https://www.readbyqxmd.com/read/27849134/developing-and-characterization-of-single-chain-variable-fragment-scfv-antibody-against-frizzled-7-fzd7-receptor
#14
Hamid Nickho, Vahid Younesi, Leili Aghebati-Maleki, Morteza Motallebnezhad, Jafar Majidi Zolbanin, Aliakbar Movassagh Pour, Mehdi Yousefi
ABSTACT Wnt/β-catenin signaling pathway through Frizzled receptors has been shown to play a key role in both normal development and tumorigenesis. Overexpression of Wnt pathway genes, such as Fzd7 in several malignancies is well-documented. Therefore, targeting of Fzd7 and its ligand inhibits cancer cells proliferation metastasis. In the present study we isolated single chain variable fragments (scFvs) against Fzd7 receptor using phage display method. Semi-synthetic human naive antibody libraries (Tomlinson I + J) was employed in panning procedure to isolate specific scFv against specific peptide from extracellular domain of Fzd7 receptor...
November 16, 2016: Bioengineered
https://www.readbyqxmd.com/read/27829339/improved-brain-expression-of-anti-amyloid-%C3%AE-scfv-by-complexation-of-mrna-including-a-secretion-sequence-with-peg-based-block-catiomer
#15
Federico Perche, Satoshi Uchida, Hiroki Akiba, Chin-Yu Lin, Masaru Ikegami, Anjaneyulu Dirisala, Toshihiro Nakashima, Keiji Itaka, Kohei Tsumoto, Kazunori Kataoka
Amyloid β aggregates have been proposed as central effectors in Alzheimer's disease, as evidenced by several passive immunotherapies in clinical trials. We used mRNA encoding anti-amyloid β (Aβ) scFv for passive immunotherapy. These scFvs could recognize amyloid oligomers with significant affinity, delay the aggregation of amyloid β in vitro, and dissociate Aβ aggregates in vitro. Intra-cerebroventricular injection of pDNA and mRNA encoding the scFv resulted in detectable scFv levels in the brain, with higher in vivo expression from mRNA...
November 8, 2016: Current Alzheimer Research
https://www.readbyqxmd.com/read/27805609/bacterial-inner-membrane-display-for-screening-a-library-of-antibody-fragments
#16
Parisa Moghaddam-Taaheri, Svetlana P Ikonomova, Zifan Gong, Janna Q Wisniewski, Amy J Karlsson
Antibodies engineered for intracellular function must not only have affinity for their target antigen, but must also be soluble and correctly folded in the cytoplasm. Commonly used methods for the display and screening of recombinant antibody libraries do not incorporate intracellular protein folding quality control, and, thus, the antigen-binding capability and cytoplasmic folding and solubility of antibodies engineered using these methods often must be engineered separately. Here, we describe a protocol to screen a recombinant library of single-chain variable fragment (scFv) antibodies for antigen-binding and proper cytoplasmic folding simultaneously...
October 15, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27786270/recombinant-antibodies-for-specific-detection-of-clostridial-fe-fe-hydrogenases
#17
Rahul Mangayil, Matti Karp, Urpo Lamminmäki, Ville Santala
Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels...
October 27, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27762541/nanoyeast-and-other-cell-envelope-compositions-for-protein-studies-and-biosensor-applications
#18
Yadveer S Grewal, Muhammad J A Shiddiky, Stephen M Mahler, Gerard A Cangelosi, Matt Trau
Rapid progress in disease biomarker discovery has increased the need for robust detection technologies. In the past several years, the designs of many immunoaffinity reagents have focused on lowering costs and improving specificity while also promoting stability. Antibody fragments (scFvs) have long been displayed on the surface of yeast and phage libraries for selection; however, the stable production of such fragments presents challenges that hamper their widespread use in diagnostics. Membrane and cell wall proteins similarly suffer from stability problems when solubilized from their native environment...
November 16, 2016: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/27712994/identification-of-high-affinity-anti-cd16a-allotype-independent-human-antibody-domains
#19
Wei Li, Hongjia Yang, Dimiter S Dimitrov
CD16A (FcγRIIIA) is an activating receptor mostly expressed on natural killer (NK) cells and monocytes/macrophages. It can mediate antibody-dependent cell-mediated cytotoxicity (ADCC) through low-affinity interaction with human immunoglobulin G (IgG) Fc. It can also mediate cell lysis if NK cells are guided by bispecific killer cells engagers (BiKEs). BiKEs showed some success in clinical trials of cancer and are promising candidate therapeutics. However, currently reported BiKEs are based on antibody fragments (scFvs) of relatively large size...
October 3, 2016: Experimental and Molecular Pathology
https://www.readbyqxmd.com/read/27692095/screening-scfv-antibodies-against-infectious-bursal-disease-virus-by-co-expression-of-antigen-and-antibody-in-the-bacteria-display-system
#20
Xiaochen Guo, Hongxue Cao, Yunxin Wang, Yang Liu, Yanmin Chen, Nan Wang, Shan Jiang, Shengqi Zhang, Qiang Wu, Tianhe Li, Yingjie Zhang, Bing Zhou, Jiechao Yin, Deshan Li, Guiping Ren
We have previously reported an antigen and antibody co-expression (AAC) technology to demonstrate the interaction between a known antigen and antibody. To validate the co-expression system for screening antibody libraries, a single chain fragment variable(scFv)antibody library was constructed from chickens immunized with the VP2 protein of infectious bursal disease virus (IBDV). The genes of both VP2 and scFv antibodies were inserted into the pBFD-Ab-Ag vector. The co-expression library was subjected to three rounds of screening by flow cytometry (FCM) using a polyclonal antibody against VP2 through a bacteria display system...
November 1, 2016: Veterinary Immunology and Immunopathology
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