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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
#1
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
#2
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...
June 22, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28771896/dual-site-specifically-modified-antibodies-with-solid-phase-immobilized-microbial-transglutaminase
#3
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
#4
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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
#11
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
#12
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
#13
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
https://www.readbyqxmd.com/read/28444391/dual-display-phage-selection-driven-by-co-engagement-of-two-targets-by-two-different-antibody-fragments
#14
Séverine Fagète, Ledicia Botas-Perez, Irène Rossito-Borlat, Kenneth Adea, Franck Gueneau, Ulla Ravn, François Rousseau, Marie Kosco-Vilbois, Nicolas Fischer, Oliver Hartley
Antibody phage display technology has supported the emergence of numerous therapeutic antibodies. The development of bispecific antibodies, a promising new frontier in antibody therapy, could be facilitated by new phage display approaches that enable pairs of antibodies to be co-selected based on co-engagement of their respective targets. We describe such an approach, making use of two complementary leucine zipper domains that heterodimerize with high affinity. Phagemids encoding a first antibody fragment (scFv) fused to phage coat protein via the first leucine zipper are rescued in bacteria expressing a second scFv fused to the second leucine zipper as a soluble periplasmic protein, so that it is acquired by phage during assembly...
April 24, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28416651/aav-mediated-expression-of-anti-tau-scfvs-decreases-tau-accumulation-in-a-mouse-model-of-tauopathy
#15
Christina Ising, Gilbert Gallardo, Cheryl E G Leyns, Connie H Wong, Floy Stewart, Lauren J Koscal, Joseph Roh, Grace O Robinson, Javier Remolina Serrano, David M Holtzman
Tauopathies are characterized by the progressive accumulation of hyperphosphorylated, aggregated forms of tau. Our laboratory has previously demonstrated that passive immunization with an anti-tau antibody, HJ8.5, decreased accumulation of pathological tau in a human P301S tau-expressing transgenic (P301S-tg) mouse model of frontotemporal dementia/tauopathy. To investigate whether the Fc domain of HJ8.5 is required for the therapeutic effect, we engineered single-chain variable fragments (scFvs) derived from HJ8...
May 1, 2017: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/28392906/selection-of-single-chain-variable-fragments-specific-for-mycobacterium-tuberculosis-esat-6-antigen-using-ribosome-display
#16
Shahrzad Ahangarzadeh, Mojgan Bandehpour, Bahram Kazemi
OBJECTIVES: Tuberculosis (TB) is still one of the problematic infectious diseases in developing countries, especially in Iran. In the present study, we applied ribosome display technique to select single chain variable fragments (scFvs) specific for the 6-kDa early secretory antigenic target (ESAT-6) antigen of Mycobacterium tuberculosis from a mouse scFv library. MATERIALS AND METHODS: The gene encoding ESAT-6 was cloned into pET22b(+) plasmid and expressed in Escherichia coli BL21 (DE3)...
March 2017: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/28392422/the-effect-of-varying-the-peptide-linker-length-in-a-single-chain-variable-fragment-antibody-against-wogonin-glucuronide
#17
Madan Kumar Paudel, Seiichi Sakamoto, Le Van Huy, Hiroyuki Tanaka, Tomofumi Miyamoto, Satoshi Morimoto
Peptide linkers of three different lengths were constructed to join the variable regions of the heavy chain (VH) and the light chain (VL) in a single-chain variable fragment antibody (scFv) specific for wogonin glucuronide (Wgn) that has the structure VH-(GGGGS)n-VL (n=3, 5, or 7). The scFv antibodies, which were expressed in Escherichia coli, were derived from an anti-Wgn monoclonal antibody (315A). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was used to evaluate their reactivity and sensitivity, which is also used for quantitative analysis of Wgn...
April 7, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28372328/blood-based-oligomeric-and-other-protein-variant-biomarkers-to-facilitate-pre-symptomatic-diagnosis-and-staging-of-alzheimer-s-disease
#18
Stephanie M Williams, Philip Schulz, Terrone L Rosenberry, Richard J Caselli, Michael R Sierks
Oligomeric forms of amyloid-β (Aβ), tau, and TDP-43 play important roles in Alzheimer's disease (AD), and therefore are promising biomarkers. We previously generated single chain antibody fragments (scFvs) that selectively bind disease-related variants of these proteins including A4, C6T, and E1, which bind different oligomeric Aβ variants; D11C, which binds oligomeric tau; and AD-TDP1 and AD-TDP2, which bind disease related TDP-43 variants. To determine the utility of these disease-related variants as early biomarkers, we first analyzed 11 human sera samples obtained ∼2 years prior to an initial mild cognitive impairment (MCI) diagnosis...
2017: Journal of Alzheimer's Disease: JAD
https://www.readbyqxmd.com/read/28359585/imaging-translational-and-post-translational-gene-regulatory-dynamics-in-living-cells-with-antibody-based-probes
#19
REVIEW
Kenneth Lyon, Timothy J Stasevich
Antibody derivatives, such as antibody fragments (Fabs) and single-chain variable fragments (scFvs), are now being used to image traditionally hard-to-see protein subpopulations, including nascent polypeptides being translated and post-translationally modified proteins. This has allowed researchers to directly image and quantify, for the first time, translation initiation and elongation kinetics with single-transcript resolution and the temporal ordering and kinetics of post-translational histone and RNA polymerase II modifications...
May 2017: Trends in Genetics: TIG
https://www.readbyqxmd.com/read/28346811/improved-soluble-scfv-elisa-screening-approach-for-antibody-discovery-using-phage-display-technology
#20
Mohammad R Tohidkia, Maryam Sepehri, Shirin Khajeh, Jaleh Barar, Yadollah Omidi
Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the "biopanning" process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs...
March 1, 2017: SLAS Discovery
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