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https://www.readbyqxmd.com/read/28900185/anti-a%C3%AE-single-chain-variable-fragment-antibodies-restore-memory-acquisition-in-a-drosophila-model-of-alzheimer-s-disease
#1
Alfonso Martin-Peña, Diego E Rincon-Limas, Pedro Fernandez-Funez
Alzheimer's disease (AD) is a prevalent neurodegenerative disorder triggered by the accumulation of soluble assemblies of the amyloid-β42 (Aβ42) peptide. Despite remarkable advances in understanding the pathogenesis of AD, the development of palliative therapies is still lacking. Engineered anti-Aβ42 antibodies are a promising strategy to stall the progression of the disease. Single-chain variable fragment (scFv) antibodies increase brain penetration and offer flexible options for delivery while maintaining the epitope targeting of full antibodies...
September 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28888872/the-catalytic-activity-of-a-recombinant-single-chain-variable-fragment-nucleic-acid-hydrolysing-antibody-varies-with-fusion-tag-and-expression-host
#2
Joungmin Lee, Minjae Kim, Youngsil Seo, Yeonjin Lee, Hyunjoon Park, Sung June Byun, Myung-Hee Kwon
The antigen-binding properties of single chain Fv antibodies (scFvs) can vary depending on the position and type of fusion tag used, as well as the host cells used for expression. The issue is even more complicated with a catalytic scFv antibody that binds and hydrolyses a specific antigen. Herein, we investigated the antigen-binding and -hydrolysing activities of the catalytic anti-nucleic acid antibody 3D8 scFv expressed in Escherichia coli or HEK293f cells with or without additional amino acid residues at the N- and C-termini...
September 6, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28887121/discovery-of-human-scfvs-that-cross-neutralize-the-toxic-effects-of-b-jararacussu-and-c-d-terrificus-venoms
#3
Luciano C Silva, Manuela B Pucca, Gabriela Pessenda, Lucas B Campos, Edson Z Martinez, Felipe A Cerni, José E Barbosa
Accidents involving venomous snakes are a public health problem worldwide, causing a large number of deaths per year. In Brazil, the majority of accidents are caused by the Bothrops and Crotalus genera, which are responsible for approximately 80% of severe envenoming cases. The cross-neutralization of snake venoms by antibodies is an important issue for development of more effective treatments. Our group has previously reported the construction of human monoclonal antibody fragments towards Bothrops jararacussu and Crotalus durissus terrificus' venoms...
September 5, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28860479/de-novo-design-of-antibody-complementarity-determining-regions-binding-a-flag-tetra-peptide
#4
Kevin C Entzminger, Jeong-Min Hyun, Robert J Pantazes, Athena C Patterson-Orazem, Ahlam N Qerqez, Zach P Frye, Randall A Hughes, Andrew D Ellington, Raquel L Lieberman, Costas D Maranas, Jennifer A Maynard
Computational antibody engineering efforts to date have focused on improving binding affinities or biophysical characteristics. De novo design of antibodies binding specific epitopes could greatly accelerate discovery of therapeutics as compared to conventional immunization or synthetic library selection strategies. Here, we employed de novo complementarity determining region (CDR) design to engineer targeted antibody-antigen interactions using previously described in silico methods. CDRs predicted to bind the minimal FLAG peptide (Asp-Tyr-Lys-Asp) were grafted onto a single-chain variable fragment (scFv) acceptor framework...
August 31, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28846502/rare-high-affinity-anti-pathogen-antibodies-from-human-repertoires-discovered-using-microfluidics-and-molecular-genomics
#5
Adam S Adler, Rena A Mizrahi, Matthew J Spindler, Matthew S Adams, Michael A Asensio, Robert C Edgar, Jackson Leong, Renee Leong, Lucy Roalfe, Rebecca White, David Goldblatt, David S Johnson
Affinity-matured, functional anti-pathogen antibodies are present at low frequencies in natural human repertoires. These antibodies are often excellent candidates for therapeutic monoclonal antibodies. However, mining natural human antibody repertoires is a challenge. In this study, we demonstrate a new method that uses microfluidics, yeast display, and deep sequencing to identify 247 natively paired anti-pathogen single-chain variable fragments (scFvs), which were initially as rare as 1 in 100,000 in the human repertoires...
August 28, 2017: MAbs
https://www.readbyqxmd.com/read/28846130/identification-and-characterization-of-the-proteins-bound-by-specific-phage-displayed-recombinant-antibodies-scfv-obtained-against-brazil-nut-and-almond-extracts
#6
Silvia de la Cruz, Raquel Madrid, Aina García, Marcos Alcocer, Rosario Martín, Isabel González, Teresa García
BACKGROUND: Almonds and Brazil nuts are widely consumed allergenic nuts whose presence must be declared according to food labelling regulations. Their detection in food products has been recently achieved by ELISA methods with recombinant antibodies (scFv) isolated against complete Brazil nut and almond protein extracts. The screening of phage-scFv libraries against complete protein extracts confers a series of advantages over the use of purified proteins, as recombinant proteins might alter their native folding...
August 28, 2017: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/28825979/covalent-coupling-of-high-affinity-ligands-to-the-surface-of-viral-vector-particles-by-protein-trans-splicing-mediates-cell-type-specific-gene-transfer
#7
Alexander Muik, Johanna Reul, Thorsten Friedel, Anke Muth, Karen Patricia Hartmann, Irene C Schneider, Robert C Münch, Christian J Buchholz
We have established a novel approach for the covalent coupling of large polypeptides to the surface of fully assembled adeno-associated viral gene transfer vector (AAV) particles via split-intein mediated protein-trans-splicing (PTS). This way, we achieved selective gene transfer to distinct cell types. Single-chain variable fragments (scFvs) or designed ankyrin repeat proteins (DARPins), exhibiting high-affinity binding to cell surface receptors selectively expressed on the surface of target cells, were coupled to AAV particles harboring mutations in the capsid proteins which ablate natural receptor usage...
July 25, 2017: Biomaterials
https://www.readbyqxmd.com/read/28807568/function-of-novel-anti-cd19-chimeric-antigen-receptors-with-human-variable-regions-is-affected-by-hinge-and-transmembrane-domains
#8
Leah Alabanza, Melissa Pegues, Claudia Geldres, Victoria Shi, Jed J W Wiltzius, Stuart A Sievers, Shicheng Yang, James N Kochenderfer
Anti-CD19 chimeric antigen receptor (CAR) T cells have caused remissions of B cell malignancies, but problems including cytokine-mediated toxicity and short persistence of CAR T cells in vivo might limit the effectiveness of anti-CD19 CAR T cells. Anti-CD19 CARs that have been tested clinically had single-chain variable fragments (scFvs) derived from murine antibodies. We have designed and constructed novel anti-CD19 CARs containing a scFv with fully human variable regions. T cells expressing these CARs specifically recognized CD19(+) target cells and carried out functions including degranulation, cytokine release, and proliferation...
July 27, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28796529/designed-ankyrin-repeat-proteins-as-her2-targeting-domains-in-chimeric-antigen-receptor-engineered-t-cells
#9
Elizabeth Siegler, Si Li, Yu Jeong Kim, Pin Wang
Chimeric antigen receptor (CAR) engineering is a branch of cancer immunotherapy that equips immune cells to target tumor antigens expressed on the cell surface using antibody-derived single-chain variable fragments (scFvs). However, other antibody mimetics, such as designed ankyrin repeat proteins (DARPins), can also serve as antigen-binding domains in CARs. This study shows that CAR-engineered T (CAR-T) cells utilizing Her2-targeting DARPins G3 and 929 can target human epidermal growth factor receptor 2 (Her2)-overexpressing cancer cells as effectively as CAR-T cells with the scFv 4D5 in vitro, and G3 CAR-T cells can slow or eliminate tumor growth in vivo as effectively as 4D5 CAR-T cells...
September 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28771896/dual-site-specifically-modified-antibodies-with-solid-phase-immobilized-microbial-transglutaminase
#10
Philipp Spycher, Christian A Amann, Jöri Wehrmüller, David R Hurwitz, Oliver Kreis, Andreas Ritler, Daniel Messmer, Andreas Küchler, Alain Blanc, Peter Walde, Martin Béhé, Roger Schibli
Microbial transglutaminase (MTG) has been stably solid-phase immobilized on glass microbeads using a second generation dendronized polymer. Immobilized MTG enabled the efficient generation of site-specifically conjugated proteins including scFvs and Fab-fragments as well as whole antibodies via distinct glutamines and unprecedented, also via lysines with various bifunctional substrates with defined stoichiometry. With this method we generated dual site-specifically modified antibodies comprising a fluorescent probe and a metal chelator for radiolabeling - a strategy anticipated to design antibodies for imaging and therapy...
August 3, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28740407/comparison-of-a-mouse-and-a-novel-human-scfv-snap-auristatin-f-drug-conjugate-with-potent-activity-against-egfr-overexpressing-human-solid-tumor-cells
#11
Mira Woitok, Diana Klose, Stefano Di Fiore, Wolfgang Richter, Christoph Stein, Gerrit Gresch, Elena Grieger, Stefan Barth, Rainer Fischer, Katharina Kolberg, Judith Niesen
Antibody-drug conjugates (ADCs) can deliver toxins to specific targets such as tumor cells. They have shown promise in preclinical/clinical development but feature stoichiometrically undefined chemical linkages, and those based on full-size antibodies achieve only limited tumor penetration. SNAP-tag technology can overcome these challenges by conjugating benzylguanine-modified toxins to single-chain fragment variables (scFvs) with 1:1 stoichiometry while preserving antigen binding. Two (human and mouse) scFv-SNAP fusion proteins recognizing the epidermal growth factor receptor (EGFR) were expressed in HEK 293T cells...
2017: OncoTargets and Therapy
https://www.readbyqxmd.com/read/28670737/a-human-scfv-antibody-that-targets-and-neutralizes-high-molecular-weight-pathogenic-amyloid-%C3%AE-oligomers
#12
Adriano Sebollela, Erika N Cline, Izolda Popova, Kevin Luo, Xiaoxia Sun, Jay Ahn, Milena A Barcelos, Vanessa N Bezerra, Natalia M Lyra E Silva, Jason Patel, Nathalia R Pinheiro, Lei A Qin, Josette M Kamel, Anthea Weng, Nadia DiNunno, Adrian M Bebenek, Pauline T Velasco, Kirsten L Viola, Pascale N Lacor, Sergio T Ferreira, William L Klein
Brain accumulation of soluble oligomers of the amyloid-β peptide (AβOs) is increasingly considered a key early event in the pathogenesis of Alzheimer's disease (AD). A variety of AβO species have been identified, both in vitro and in vivo, ranging from dimers to 24mers and higher-order oligomers. However, there is no consensus in the literature regarding which AβO species are most germane to AD pathogenesis. Antibodies capable of specifically recognizing defined subpopulations of AβOs would be a valuable asset in the identification, isolation, and characterization of AD-relevant AβO species...
July 3, 2017: Journal of Neurochemistry
https://www.readbyqxmd.com/read/28641985/single-step-colony-assay-for-screening-antibody-libraries
#13
Mieko Kato, Yoshiro Hanyu
We describe a method, single-step colony assay, for simple and rapid screening of single-chain Fv fragment (scFv) libraries. Colonies of Escherichia coli expressing the scFv library are formed on a hydrophilic filter that is positioned in contact with a membrane coated with an antigen. scFv expression is triggered upon treatment of colonies with an induction reagent, following which scFvs are secreted from the cells and diffused to the antigen-coated membrane. scFvs that exhibit binding affinity for the antigen are captured by the membrane-immobilized antigen...
June 20, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28621534/construction-of-an-immunized-rabbit-phage-display-library-for-selecting-high-activity-against-bacillus-thuringiensis-cry1f-toxin-single-chain-antibodies
#14
Chongxin Xu, Cunzheng Zhang, Jianfeng Zhong, Hui Hu, Shimin Luo, Xiaoqin Liu, Xiao Zhang, Yuan Liu, Xianjin Liu
In the present study, a Cry1F-immunized rabbit phage display library (6.96 × 10(8) cfu/mL) was constructed for selecting high activity of anti-Cry1F toxin single-chain antibody (a single-chain variable fragment, scFv) by biopanning. A total of 16 positive monoclonal phage scFv's were obtained after 4 rounds of panning, which were identified by enzyme-linked immunosorbent assay (ELISA), polymerized chain reaction, and DNA sequencing. The most positive phage scFv (named RF4) was expressed in Escherichia coli HB2151, and a soluble protein of approximately 30 kDa was purified with sodium dodecyl sulfate-polyacrylamide gel electrophoresis...
July 26, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28591728/generation-and-evaluation-of-antibody-agents-for-molecular-imaging-of-cd44v6-expressing-cancers
#15
Anna-Karin Haylock, Johan Nilvebrant, Anja Mortensen, Irina Velikyan, Marika Nestor, Ronny Falk
AIM: The aim of this study was to generate and characterize scFv antibodies directed to human CD44v6, as well as to radiolabel and evaluate top candidates in vitro and in vivo for their potential use in CD44v6-targeted molecular imaging in cancer patients. MATERIALS AND METHODS: Phage display selections were used to isolate CD44v6-specific scFvs. A chain shuffling strategy was employed for affinity maturation based on a set of CD44v6-specific first-generation clones...
May 18, 2017: Oncotarget
https://www.readbyqxmd.com/read/28584111/phage-display-and-kinetic-selection-of-antibodies-that-specifically-inhibit-amyloid-self-replication
#16
Anna Munke, Jonas Persson, Tanja Weiffert, Erwin De Genst, Georg Meisl, Paolo Arosio, Anna Carnerup, Christopher M Dobson, Michele Vendruscolo, Tuomas P J Knowles, Sara Linse
The aggregation of the amyloid β peptide (Aβ) into amyloid fibrils is a defining characteristic of Alzheimer's disease. Because of the complexity of this aggregation process, effective therapeutic inhibitors will need to target the specific microscopic steps that lead to the production of neurotoxic species. We introduce a strategy for generating fibril-specific antibodies that selectively suppress fibril-dependent secondary nucleation of the 42-residue form of Aβ (Aβ42). We target this step because it has been shown to produce the majority of neurotoxic species during aggregation of Aβ42...
June 20, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28544767/a-poly-propyleneimine-dendrimer-based-polyplex-system-for-single-chain-antibody-mediated-targeted-delivery-and-cellular-uptake-of-sirna
#17
Stefanie Tietze, Isabell Schau, Susanne Michen, Franka Ennen, Andreas Janke, Gabriele Schackert, Achim Aigner, Dietmar Appelhans, Achim Temme
Therapeutics based on small interfering RNAs (siRNAs) offer a great potential to treat so far incurable diseases or metastatic cancer. However, the broad application of siRNAs using various nonviral carrier systems is hampered by unspecific toxic side effects, poor pharmacokinetics due to unwanted delivery of siRNA-loaded nanoparticles into nontarget organs, or rapid renal excretion. In order to overcome these obstacles, several targeting strategies using chemically linked antibodies and ligands have emerged...
May 22, 2017: Small
https://www.readbyqxmd.com/read/28536241/correction-aav-mediated-expression-of-anti-tau-scfvs-decreases-tau-accumulation-in-a-mouse-model-of-tauopathy
#18
Christina Ising, Gilbert Gallardo, Cheryl E G Leyns, Connie H Wong, Hong Jiang, Floy Stewart, Lauren J Koscal, Joseph Roh, Grace O Robinson, Javier Remolina Serrano, David M Holtzman
No abstract text is available yet for this article.
July 3, 2017: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/28491282/cell-growth-inhibition-and-apoptotic-effects-of-a-specific-anti-rtfscfv-antibody-on-prostate-cancer-but-not-glioblastoma-cells
#19
Foroogh Nejatollahi, Payam Bayat, Bahareh Moazen
Background: Single chain antibody (scFv) has shown interesting results in cancer immunotargeting approaches, due to its advantages over monoclonal antibodies. Regeneration and tolerance factor (RTF) is one of the most important regulators of extracellular and intracellular pH in eukaryotic cells. In this study, the inhibitory effects of a specific anti-RTF scFv were investigated and compared between three types of prostate cancer and two types of glioblastoma cells.  Methods: A phage antibody display library of scFv was used to select specific scFvs against RTF using panning process...
2017: F1000Research
https://www.readbyqxmd.com/read/28445413/identification-of-highly-specific-scfvs-against-total-adiponectin-for-diagnostic-purposes
#20
Peter Wilton, Michael Steidel, Gabriele Krczal, Iris Hermanns, Andreas Pfützner, Alisa Konnerth, Kajohn Boonrod
Adiponectin is one of the most abundant adipokines secreted from adipose tissue. It acts as an endogenous insulin sensitizer and plasma concentrations are inversely correlated with obesity and metabolic syndrome. A decrease in plasma adiponectin levels normally indicates increased hormonal activity of the visceral lipid tissue, which is associated with decreased insulin sensitivity. It may therefore be considered a valuable biomarker for elucidating the underlying deteriorations resulting in type 2 diabetes and macrovascular disease...
April 26, 2017: Biology
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