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https://www.readbyqxmd.com/read/28724948/transgenic-chickens-expressing-the-3d8-single-chain-variable-fragment-protein-suppress-avian-influenza-transmission
#1
Sung June Byun, Seong-Su Yuk, Ye-Jin Jang, Hoonsung Choi, Mi-Hyang Jeon, T O Erdene-Ochir, Jung-Hoon Kwon, Jin-Yong Noh, Jeom Sun Kim, Jae Gyu Yoo, Chang-Seon Song
The 3D8 single chain variable fragment (scFv) is a mini-antibody that causes unusual sequence-independent nuclease activity against all types of nucleic acids. We used recombinant lentiviruses to generate transgenic chickens expressing the 3D8 scFv gene under the control of the chicken β-actin promoter. From 420 injected embryos, 200 chicks (G0) hatched and were screened for the 3D8 scFv using PCR, and 15 chicks were identified as transgenic birds expressing the transgene in their semen. The G0 founder birds were mated with wild-type hens to produce seven transgenic chicks (G1)...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28708284/cd20-specific-immunoligands-engaging-nkg2d-enhance-%C3%AE-%C3%AE-t-cell-mediated-lysis-of-lymphoma-cells
#2
Matthias Peipp, Daniela Wesch, Hans-Heinrich Oberg, Sebastian Lutz, Anja Muskulus, Jan G J van de Winkel, Paul W H I Parren, Renate Burger, Andreas Humpe, Dieter Kabelitz, Martin Gramatzki, Christian Kellner
Human γδ T cells are innate-like T cells which are able to kill a broad range of tumor cells and thus may have potential for cancer immunotherapy. The activating receptor natural killer group 2 member D (NKG2D) plays a key role in regulating immune responses driven by γδ T cells. Here, we explored whether recombinant immunoligands consisting of a CD20 single-chain fragment variable (scFv) linked to a NKG2D ligand, either MHC class I chain-related protein A (MICA) or UL16 binding protein 2 (ULBP2), could be employed to engage γδ T cells for tumor cell killing...
July 14, 2017: Scandinavian Journal of Immunology
https://www.readbyqxmd.com/read/28707665/-human-selenium-containing-single-chain-variable-fragment-with-glutathione-peroxidase-activity-protects-nih3t3-fibroblast-against-oxidative-damage
#3
R Huo, L Yang, T G Zhang, J Y Wei
Ultraviolet B (UVB medium wave, 280-315 nm) induces cellular oxidative damage and apoptosis by producing reactive oxygen species (ROS). Glutathione peroxidase functions as an antioxidant by catalyzing the reduction of hydrogen peroxide, the more important member of reactive oxygen species. A human selenium-containing single-chain variable fragment (se-scFv-B3) with glutathione peroxidase activity of 1288 U/μmol was generated and investigated for its antioxidant effects in UVB-induced oxidative damage model...
May 2017: Molekuliarnaia Biologiia
https://www.readbyqxmd.com/read/28699800/towards-the-improvement-in-stability-of-an-anti-a%C3%AE-single-chain-variable-fragment-scfv-h3d6-as-a-way-to-enhance-its-therapeutic-potential
#4
Laia Montoliu-Gaya, Javier Murciano-Calles, Jose C Martinez, Sandra Villegas
ScFv-h3D6 is a single-chain variable fragment derived from the monoclonal antibody bapineuzumab that prevents Aβ-induced cytotoxicity by capturing Aβ oligomers. The benefits of scFv-h3D6 treatment in Alzheimer's disease are known at the behavioural, cellular and molecular levels in the 3xTg-AD mouse model. Antibody-based therapeutics are only stable in a limited temperature range, so their benefit in vivo depends on their capability for maintaining the proper fold. Here, we have stabilized the scFv-h3D6 folding by introducing the mutation VH-K64R and combining it with the previously described elongation of the VL domain (C3)...
July 12, 2017: Amyloid: the International Journal of Experimental and Clinical Investigation
https://www.readbyqxmd.com/read/28699736/detection-of-an-integrin-binding-mechanoswitch-within-fibronectin-during-tissue-formation-and-fibrosis
#5
Lizhi Cao, John Nicosia, Jacqueline Larouche, Yuanyuan Zhang, Haylee Bachman, Ashley C Brown, Lars Holmgren, Thomas H Barker
Fibronectin (Fn) is an extracellular matrix protein that orchestrates complex cell adhesion and signaling through cell surface integrin receptors during tissue development, remodeling, and disease, such as fibrosis. Fn is sensitive to mechanical forces in its tandem type III repeats, resulting in extensive molecular enlongation. As such, it has long been hypothesized that cell- and tissue-derived forces may activate an "integrin switch" within the critical integrin-binding ninth and 10th type III repeats-conferring differential integrin-binding specificity, leading to differential cell responses...
July 12, 2017: ACS Nano
https://www.readbyqxmd.com/read/28692735/an-anti-vegf-b-antibody-fragment-induces-regression-of-pre-existing-blood-vessels-in-the-rat-cornea
#6
Yazad D Irani, Pierre D Scotney, Sonja Klebe, Lauren A Mortimer, Andrew D Nash, Keryn A Williams
Purpose: We tested the ability of an antibody fragment with specificity for vascular endothelial growth factor-B (VEGF-B) to regress nascent and established corneal blood vessels in the rat. Methods: A single chain variable antibody fragment (scFv) with specificity for VEGF-B was engineered from the 2H10 hybridoma. Binding to rat, mouse, and human VEGF-B was confirmed by surface plasmon resonance. Activity of the anti-VEGF-B scFv on developing and established corneal blood vessels was assessed following unilateral superficial cautery in male and female outbred Sprague Dawley rats...
July 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28689385/expression-optimization-of-anti-cd22-scfv-apoptin-fusion-protein-using-experimental-design-methodology
#7
Solmaz Agha Amiri, Najmeh Zarei, Somayeh Enayati, Mohammad Azizi, Vahid Khalaj, Soraya Shahhosseini
Background: Design of experiments is a rapid and cost-effective approach for optimization of recombinant protein production process. In our previous study, we generated a potent dual-acting fusion protein, anti-CD22 scFv-apoptin, to target B-cell malignant cell lines. In the present investigation, we report the effect of different variables on the expression levels of this fusion protein. Methods: : Four variables (cell optical density at induction, IPTG concentration, induction temperature, and induction time) were tested using experimental design...
July 10, 2017: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/28680755/bispecific-antibody-does-not-induce-t-cell-death-mediated-by-chimeric-antigen-receptor-against-disialoganglioside-gd2
#8
Sayed Shahabuddin Hoseini, Konstantin Dobrenkov, Dmitry Pankov, Xiaoliang L Xu, Nai-Kong V Cheung
Chimeric antigen receptors (CAR) and bispecific antibodies (BsAb) are two powerful immunotherapy approaches for retargeting lymphocytes toward cancer cells. Despite their success in lymphoblastic leukemia, solid tumors have been more recalcitrant. Identifying therapeutic barriers facing CAR-modified (CART) or BsAb-redirected T (BsAb-T) cells should facilitate their clinical translation to solid tumors. Novel lentiviral vectors containing low-affinity or high-affinity 4-1BB second-generation anti-GD2 (disialoganglioside) CARs were built to achieve efficient T cell transduction...
2017: Oncoimmunology
https://www.readbyqxmd.com/read/28674295/development-of-a-rapid-identification-method-for-a-variety-of-antibody-candidates-using-high-throughput-sequencing
#9
Yuji Ito
 As an alternative to hybridoma technology, the antibody phage library system can also be used for antibody selection. This method enables the isolation of antigen-specific binders through an in vitro selection process known as biopanning. While it has several advantages, such as an avoidance of animal immunization, the phage cloning and screening steps of biopanning are time-consuming and problematic. Here, we introduce a novel biopanning method combined with high-throughput sequencing (HTS) using a next-generation sequencer (NGS) to save time and effort in antibody selection, and to increase the diversity of acquired antibody sequences...
2017: Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan
https://www.readbyqxmd.com/read/28670737/a-human-scfv-antibody-that-targets-and-neutralizes-high-molecular-weight-pathogenic-amyloid-%C3%AE-oligomers
#10
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/28669727/light-chain-residue-95-is-critical-for-antigen-binding-and-multispecificity-of-monoclonal-antibody-g2
#11
Daiki Usui, Satomi Inaba, Yuji O Kamatari, Naotaka Ishiguro, Masayuki Oda
The monoclonal antibody, G2, specifically binds to the immunogen peptide derived from the chicken prion protein, Pep18mer, and two chicken proteins derived peptides, Pep8 and Pep395; G2 binds with equal affinity to Pep18mer. The amino acid sequences of the three peptides are completely different, and so the recognition mechanism of G2 is unique and interesting. We generated a single-chain Fv (scFv) antibody of G2, and demonstrated its correct folding with an antigen binding function similar to intact G2 antibody...
June 29, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28669053/efficient-targeting-of-cd13-on-cancer-cells-by-the-immunotoxin-scfv13-eta-and-the-bispecific-scfv-13xds16
#12
Elena Grieger, Gerrit Gresch, Judith Niesen, Mira Woitok, Stefan Barth, Rainer Fischer, Rolf Fendel, Christoph Stein
PURPOSE: Treatment of cancer using standard chemotherapy still offers a poor prognosis combined with severe side effects. Novel antibody-based therapies have been shown to overcome low efficiency and lack of selectivity by targeting cancer-associated antigens, such as aminopeptidase CD13. METHODS: We isolated a high-affinity CD13-specific single-chain fragment variable (scFv13) from a phage display library of V-genes from mice immunized with soluble antigen. An immunotoxin comprising the scFv13 and a truncated version of the exotoxin A of Pseudomonas aeruginosa (ETA', scFv13-ETA') and a bispecific scFv targeting CD13 and CD16 simultaneously (bsscFv[13xds16]) was generated and investigated for their therapeutic potential...
July 1, 2017: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/28667390/novel-psca-targeting-scfv-fusion-proteins-for-diagnosis-and-immunotherapy-of-prostate-cancer
#13
Claudia Kessler, Alessa Pardo, Mehmet K Tur, Stefan Gattenlöhner, Rainer Fischer, Katharina Kolberg, Stefan Barth
PURPOSE: Despite great progress in the diagnosis and treatment of localized prostate cancer (PCa), there remains a need for new diagnostic markers that can accurately distinguish indolent and aggressive variants. One promising approach is the antibody-based targeting of prostate stem cell antigen (PSCA), which is frequently overexpressed in PCa. Here, we show the construction of a molecular imaging probe comprising a humanized scFv fragment recognizing PSCA genetically fused to an engineered version of the human DNA repair enzyme O6-alkylguanine-DNA alkyltransferase (AGT), the SNAP-tag, enabling specific covalent coupling to various fluorophores for diagnosis of PCa...
June 30, 2017: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/28662917/accumulative-scfv-fc-antibody-gene-integration-into-the-hprt-chromosomal-locus-of-chinese-hamster-ovary-cells
#14
Xue Wang, Yoshinori Kawabe, Risa Kato, Takeshi Hada, Akira Ito, Yoshimasa Yamana, Masako Kondo, Masamichi Kamihira
We have previously developed an accumulative site-specific gene integration system (AGIS) using Cre-recombinase and mutated loxP sites. AGIS enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. However, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. In the present study, we describe an improved version of AGIS that facilitates and accelerates the establishment of high-producer Chinese hamster ovary (CHO) cells...
June 26, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28654662/generation-of-a-rabbit-single-chain-fragment-variable-scfv-antibody-for-specific-detection-of-bradyrhizobium-sp-doa9-in-both-free-living-and-bacteroid-forms
#15
Nguyen Xuan Vu, Natcha Pruksametanan, Witsanu Srila, Watcharin Yuttavanichakul, Kamonluck Teamtisong, Neung Teaumroong, Nantakorn Boonkerd, Panlada Tittabutr, Montarop Yamabhai
A simple and reliable method for the detection of specific nitrogen-fixing bacteria in both free-living and bacteroid forms is essential for the development and application of biofertilizer. Traditionally, a polyclonal antibody generated from an immunized rabbit was used for detection. However, the disadvantages of using a polyclonal antibody include limited supply and cross-reactivity to related bacterial strains. This is the first report on the application of phage display technology for the generation of a rabbit recombinant monoclonal antibody for specific detection and monitoring of nitrogen-fixing bacteria in both free-living form and in plant nodules...
2017: PloS One
https://www.readbyqxmd.com/read/28651553/ig-and-tr-single-chain-fragment-variable-scfv-sequence-analysis-a-new-advanced-functionality-of-imgt-v-quest-and-imgt-highv-quest
#16
Véronique Giudicelli, Patrice Duroux, Sofia Kossida, Marie-Paule Lefranc
BACKGROUND: IMGT®, the international ImMunoGeneTics information system® ( http://www.imgt.org ), was created in 1989 in Montpellier, France (CNRS and Montpellier University) to manage the huge and complex diversity of the antigen receptors, and is at the origin of immunoinformatics, a science at the interface between immunogenetics and bioinformatics. Immunoglobulins (IG) or antibodies and T cell receptors (TR) are managed and described in the IMGT® databases and tools at the level of receptor, chain and domain...
June 26, 2017: BMC Immunology
https://www.readbyqxmd.com/read/28643153/a-3e8-scfv-cys-ir800-conjugate-targeting-tag-72-in-an-orthotopic-colorectal-cancer-model
#17
Li Gong, Haiming Ding, Nicholas E Long, Brandon J Sullivan, Edward W Martin, Thomas J Magliery, Michael F Tweedle
PURPOSE: Optical surgical navigation (OSN) will be a potent tool to help surgeons more accurately and efficiently remove tumors. The purpose of this study was to evaluate a novel humanized 3E8 antibody (3E8 MAb) fragment site-specifically conjugated with IR800, 3E8.scFv.Cys-IR800, as a potential OSN agent to target colorectal adenocarcinoma. PROCEDURES: An engineered single-chain variable fragment of 3E8 MAb (targeted to TAG-72), appending a C-terminal cysteine residue (3E8...
June 22, 2017: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
https://www.readbyqxmd.com/read/28641985/single-step-colony-assay-for-screening-antibody-libraries
#18
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/28640942/polyfunctional-response-by-immtac-imcgp100-redirected-cd8-and-cd4-t-cells
#19
Caroline Boudousquie, Giovanna Bossi, Jacob M Hurst, Karolina A Rygiel, Bent K Jakobsen, Namir J Hassan
The success of immune system based cancer therapies depends on a broad immune response engaging a range of effector cells and mechanisms. Immune mobilising monoclonal TCRs against cancer (ImmTAC(™) molecules, fusion proteins consisting of a soluble, affinity enhanced TCR and an anti-CD3 scFv Ab) were previously shown to redirect CD8(+) and CD4(+) T cells against tumours. Here we present evidence that IMCgp100 (ImmTAC recognising a peptide derived from the melanoma-specific protein, gp100, presented by HLA-A*0201) efficiently redirects and activates effector and memory cells from both CD8(+) and CD4(+) repertoires...
June 22, 2017: Immunology
https://www.readbyqxmd.com/read/28621534/construction-of-an-immunized-rabbit-phage-display-library-for-selecting-high-activity-of-against-bacillus-thuringiensis-cry1f-toxin-single-chain-antibodies
#20
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 (6.96×108) was constructed for selecting high activity of anti-Cry1F toxin single-chain antibody (scFv) by bio-panning. Total 16 positive monoclonal phage scFvs were obtained after 4 rounds panning which identified by enzyme-linked immunoassay (ELISA), PCR and DNA sequencing. The most positive phage scFv (named RF4) was expressed in E.coli.HB2151 and the purified soluble protein approximate 30 kDa with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)...
June 16, 2017: Journal of Agricultural and Food Chemistry
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