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https://www.readbyqxmd.com/read/29689715/antibody-fusion-proteins-with-human-ribonucleases-1-to-8
#1
Xenia Wezler, Stefan Dübel, Thomas Schirrmann
ImmunoRNases combine tumor targeting by antibodies with the cytotoxic action of ribonucleases from the RNase A superfamily. This study investigated for the first time all catalytic active human RNase A family members (1 to 8) as effector components of antibody fusion proteins. ImmunoRNase fusion proteins were constructed using the CD30-specific bivalent recombinant scFv-Fc antibody SH313-B5. Production of the resulting entirely human immunoRNases 1 to 8 was done in mammalian cells by secretion of active forms...
April 10, 2018: Human Antibodies
https://www.readbyqxmd.com/read/29686683/high-affinity-ligands-can-trigger-t-cell-receptor-signaling-without-cd45-segregation
#2
Mohammad Ameen Al-Aghbar, Yeh-Shiu Chu, Bing-Mae Chen, Steve R Roffler
How T cell receptors (TCRs) are triggered to start signaling is still not fully understood. It has been proposed that segregation of the large membrane tyrosine phosphatase CD45 from engaged TCRs initiates signaling by favoring phosphorylation of immunoreceptor tyrosine-based activation motifs (ITAMs) in the cytoplasmic domains of CD3 molecules. However, whether CD45 segregation is important to initiate triggering is still uncertain. We examined CD45 segregation from TCRs engaged to anti-CD3 scFv with high or low affinity and with defined molecular lengths on glass-supported lipid bilayers using total internal reflection microscopy...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29678657/development-and-evaluation-of-an-optimal-human-single-chain-variable-fragment-derived-bcma-targeted-car-t-cell-vector
#3
Eric L Smith, Mette Staehr, Reed Masakayan, Ishan J Tatake, Terence J Purdon, Xiuyan Wang, Pei Wang, Hong Liu, Yiyang Xu, Sarah C Garrett-Thomson, Steven C Almo, Isabelle Riviere, Cheng Liu, Renier J Brentjens
B cell maturation antigen (BCMA) has recently been identified as an important multiple myeloma (MM)-specific target for chimeric antigen receptor (CAR) T cell therapy. In CAR T cell therapy targeting CD19 for lymphoma, host immune anti-murine CAR responses limited the efficacy of repeat dosing and possibly long-term persistence. This clinically relevant concern can be addressed by generating a CAR incorporating a human single-chain variable fragment (scFv). We screened a human B cell-derived scFv phage display library and identified a panel of BCMA-specific clones from which human CARs were engineered...
March 28, 2018: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/29670605/antiviral-immunotoxin-against-bovine-herpesvirus-1-targeted-inhibition-of-viral-replication-and-apoptosis-of-infected-cell
#4
Jian Xu, Xiaoyang Li, Bo Jiang, Xiaoyu Feng, Jing Wu, Yunhong Cai, Xixi Zhang, Xiufen Huang, Joshua E Sealy, Munir Iqbal, Yongqing Li
Bovine herpesvirus 1 (BoHV-1) is a highly contagious viral pathogen which causes infectious bovine rhinotracheitis in cattle worldwide. Currently, there is no antiviral prophylactic treatment available capable of mitigating the disease impact and facilitating recovery from latent infection. In this study, we have engineered a novel recombinant anti-BoHV-1 immunotoxin construct termed "BoScFv-PE38" that consists of a single-chain monoclonal antibody fragment (scFv) fused with an active domain of Pseudomonas exotoxin A as a toxic effector (PE38)...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29669811/linker-engineering-in-anti-tag-72-antibody-fragments-optimizes-biophysical-properties-serum-half-life-and-high-specificity-tumor-imaging
#5
Nicholas E Long, Brandon J Sullivan, Haiming Ding, Stephanie R Doll, Michael A Ryan, Charles L Hitchcock, Edward W Martin, Krishan Kumar, Michael F Tweedle, Thomas J Magliery
Antibody (Ab) fragments have great clinical potential as cancer therapeutics and diagnostics. Their small size allows for fast clearance from blood, low immunoreactivity, better tumor penetration, and simpler engineering and production. The smallest fragment derived from a full-length IgG that retains binding to its antigen, the single-chain variable fragment (scFV), is engineered by fusing the variable light and variable heavy domains with a peptide linker. Along with switching the domain orientation, altering the length and amino acid sequence of the linker can significantly affect scFV binding, stability, quaternary structure, and other biophysical properties...
April 18, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29669810/structural-insights-into-the-mechanism-of-action-of-a-biparatopic-anti-her2-antibody
#6
Vaheh Oganesyan, Li Peng, Jared S Bee, John Li, Samuel R Perry, Frank Comer, Linda Xu, Kimberly Cook, Kannaki Senthil, Lori Clarke, Kim Rosenthal, Chanshou Gao, Melissa Damschroder, Herren Wu, William F Dall'Acqua
Pathways of human EGF receptors are activated upon ligand dependent or independent homo or heterodimerization and their subsequent transphosphorylation. Overexpression of these receptors positively correlates with transphosphorylation rates and increased tumor growth rates. MEDI4276, an anti human epidermal growth factor receptor 2 (HER2) biparatopic antibody drug conjugate, has two paratopes within each of antibody arms. One, 39S, is aiming at the HER2 site involved in receptor dimerization and the second, single chain fragment (scFv) mimicking trastuzumab...
April 18, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29662239/elimination-of-tdp-43-inclusions-linked-to-amyotrophic-lateral-sclerosis-by-a-misfolding-specific-intrabody-with-dual-proteolytic-signals
#7
Yoshitaka Tamaki, Akemi Shodai, Toshifumi Morimura, Ryota Hikiami, Sumio Minamiyama, Takashi Ayaki, Ikuo Tooyama, Yoshiaki Furukawa, Ryosuke Takahashi, Makoto Urushitani
Aggregation of TAR DNA-binding protein of 43 kDa (TDP-43) is implicated in the pathogenesis of sporadic and certain familial forms of amyotrophic lateral sclerosis (ALS), suggesting elimination of TDP-43 aggregates as a possible therapeutic strategy. Here we generated and investigated a single-chain variable fragment (scFv) derived from the 3B12A monoclonal antibody (MAb) that recognises D247 of the TDP-43 nuclear export signal, an epitope masked in the physiological state. In transfected HEK293A cells, 3B12A scFv recapitulated the affinity of the full-length MAb to mislocalised TDP-43 with a defective nuclear localising signal and to a TDP-43 inclusion mimic with cysteine-to-serine substitution at RRM1...
April 16, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29651657/genetic-and-physiological-basis-for-antibody-production-by-kluyveromyces-marxianus
#8
Yumiko Nambu-Nishida, Keiji Nishida, Tomohisa Hasunuma, Akihiko Kondo
Kluyveromyces marxianus is a thermotolerant, crabtree-negative yeast, which preferentially directs metabolism (e.g., from the tricarboxylic acid cycle) to aerobic alcoholic fermentation. Thus K. marxianus has great potential for engineering to produce various materials under aerobic cultivation conditions. In this study, we engineered K. marxianus to produce and secrete a single-chain antibody (scFv), a product that is highly valuable but has historically proven difficult to generate at large scale. scFv production was obtained with strains carrying either plasmid-borne or genomically integrated constructs using various combinations of promoters (P MDH1 or P ACO1 ) and secretion signal peptides (KmINUss or Scα-MFss)...
April 12, 2018: AMB Express
https://www.readbyqxmd.com/read/29629470/biotinylated-single-chain-variable-fragment-based-enzyme-linked-immunosorbent-assay-for-glycocholic-acid
#9
Xiping Cui, Natalia Vasylieva, Ding Shen, Bogdan Barnych, Jun Yang, Qiyi He, Zhengyun Jiang, Suqing Zhao, Bruce D Hammock
Glycocholic acid (GCA) has been identified as a novel selective and sensitive biomarker for hepatocellular carcinoma (HCC). In this work, a recombinant antibody, scFv-G11, which was shown previously to have selective reactivity for GCA, was labeled with biotin using a chemical and an enzymatic method, respectively. The enzymatic method proved superior giving sensitive scFv-biotin preparations. Based on biotinylated scFv against GCA and a biotin-streptavidin system for signal amplification, an indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) has been established for the sensitive and rapid detection of GCA...
April 9, 2018: Analyst
https://www.readbyqxmd.com/read/29628793/challenging-tumour-immunological-techniques-that-help-to-track-cancer-stem-cells-in-malignant-melanomas-and-other-solid-tumours
#10
Beatrix Kotlan, Vanda Plotar, Klara Eles, Szabolcs Horvath, Timea Balatoni, Orsolya Csuka, Mihaly Újhelyi, Ákos Sávolt, Andras Szollar, Istvan Vamosi-Nagy, Laszlo Toth, Emil Farkas, Jozsef Toth, Miklos Kasler, Gabriella Liszkay
Aim of the study: The arsenal of questions and answers about the minor cancer initiating cancer stem cell (CSC) population put responsible for cancer invasiveness and metastases, has left with an unsolved puzzle. Specific aims of a complex project were partly focused on revealing new biomarkers of cancer. We designed and set up novel techniques to facilitate the detection of cancerous cells. Materials and methods: As a novel approach, we investigated B cells infiltrating breast carcinomas and melanomas (TIL-B) in terms of their tumour antigen binding potential...
March 2018: Contemporary Oncology Współczesna Onkologia
https://www.readbyqxmd.com/read/29627856/generation-and-characterization-of-a-novel-recombinant-scfv-antibody-specific-for-campylobacter-jejuni
#11
Ruramayi M Nzuma, Fuquan Liu, Irene R Grant
Campylobacter jejuni is a leading cause of foodborne illness worldwide, mainly due to consumption and handling of contaminated raw chicken. Rapid detection methods for C. jejuni are vital for monitoring contamination levels in chicken products and reducing human Campylobacteriosis cases. The 'gold standard' culture-based method of Campylobacter detection takes 3-5 days and is too slow to permit effective intervention. Immuno-based methods are faster, but usually necessitate use of animals or hybridoma technology to produce antibodies; making them difficult and expensive to produce...
April 7, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29625180/a%C3%AE-oligomer-uptake-and-the-resulting-inflammatory-response-in-adult-human-astrocytes-are-precluded-by-an-anti-a%C3%AE-single-chain-variable-fragment-in-combination-with-an-apoe-mimetic-peptide
#12
Laia Montoliu-Gaya, Sandra D Mulder, Maaike A C Herrebout, Johannes C Baayen, Sandra Villegas, Robert Veerhuis
An imbalance between production and clearance of soluble amyloid-β (Aβ) initiates the pathological process in sporadic Alzheimer's disease (AD). Aβ-specific antibodies seemed promising as therapeutic option in AD mouse models. In patients, however, vascular side-effects and Aβ-antibody complex-induced microglial and/or perivascular macrophage inflammatory responses were encountered. To prevent inflammatory reactions, we designed a single chain variable fragment (scFv-h3D6), based on monoclonal antibody bapineuzumab (mAb-h3D6), but lacking the Fc region...
April 3, 2018: Molecular and Cellular Neurosciences
https://www.readbyqxmd.com/read/29615399/human-gucy2c-targeted-chimeric-antigen-receptor-car-expressing-t-cells-eliminate-colorectal-cancer-metastases
#13
Michael S Magee, Tara S Abraham, Trevor R Baybutt, John C Flickinger, Natalie A Ridge, Glen P Marszalowicz, Priyanka Prajapati, Adam R Hersperger, Scott A Waldman, Adam E Snook
One major hurdle to the success of adoptive T-cell therapy is the identification of antigens that permit effective targeting of tumors in the absence of toxicities to essential organs. Previous work has demonstrated that T cells engineered to express chimeric antigen receptors (CAR-T cells) targeting the murine homolog of the colorectal cancer antigen GUCY2C treat established colorectal cancer metastases, without toxicity to the normal GUCY2C-expressing intestinal epithelium, reflecting structural compartmentalization of endogenous GUCY2C to apical membranes comprising the intestinal lumen...
April 3, 2018: Cancer Immunology Research
https://www.readbyqxmd.com/read/29614840/detection-of-metallothionein-in-javanese-medaka-oryzias-javanicus-using-a-scfv-immobilized-protein-chip
#14
Euiyeon Lee, Hyunjin Jeon, Chungwon Kang, Seonock Woo, Seungshic Yum, Youngeun Kwon
Environmental pollution by various industrial chemicals and biological agents poses serious risks to human health. Especially, marine contamination by potentially toxic elements (PTEs) has become a global concern in recent years. Many efforts have been undertaken to monitor the PTE contamination of the aquatic environment. However, there are few approaches available to assess the PTE exposure of aquatic organisms. In this research, we developed a strategy to evaluate the heavy metal exposure of marine organisms, by measuring the expression levels of metallothionein protein derived from Oryzias javanicus (OjaMT)...
April 2, 2018: Sensors
https://www.readbyqxmd.com/read/29609459/modulating-antibody-structure-and-function-through-directed-mutations-and-chemical-rescue
#15
Christine Kaiser, Juan Pablo Rincon Pabon, Jake Khowsathit, Paola Castaldi, Steven L Kazmirski, David D Weis, Andrew X Zhang, John Karanicolas
Monoclonal antibody therapeutics have revolutionized the treatment of diseases such as cancer and autoimmune disorders, and also serve as research reagents for diverse and unparalleled applications. To extend their utility in both contexts, we have begun development of tunable antibodies, whose activity can be controlled by addition of a small molecule. Conceptually, we envision that incorporating cavity-forming mutations into an antibody can disrupt its structure, thereby reducing its affinity for antigen; addition of a small molecule may then restore the active structure, and thus rescue antigen binding...
April 2, 2018: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29596991/a-novel-strategy-to-produce-high-level-and-high-purity-of-bioactive-il15-fusion-proteins-from-mammalian-cells
#16
Haomin Huang, Yuying Luo, Hanna Baradei, Shan Liu, Keneshia K Haenssen, Supriya Sanglikar, Senthil Kumar, John Cini
IL15, a member of the common γ chain receptor (γc) cytokine family, is gaining attention in recent years as one of the most promising anti-tumor agents. IL15 regulates T cell activation and proliferation, promotes the survival of CD8+ CD44hi memory T cells and is also essential for NK cell expansion and development. Despite the attraction of developing IL15 as an anti-cancer agent, production of recombinant IL15 has proven to be difficult due to the stringent control of IL15 expression at the transcriptional, translational and the post-translational levels...
March 26, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29575613/humanization-of-fibroblast-growth-factor-1-single-chain-antibody-and-validation-for-its-antitumorigenic-efficacy-in-breast-cancer-and-glioma-cells
#17
Xiao-Xiao He, Shuang Du, Shi-Qian Gao, Jing-Ying Chen, Ran-Juan Cao, Zhen-Kai Xing, Alia Rizvi Syeda Kazim, Hua-Li Yu, Qing-Chuan Zheng, Xiao-Juan Zhu
Single-chain variable fragment (scFv) antibodies are the smallest immunoglobulins with high antigen-binding affinity. We have previously reported that fibroblast growth factor 1 played pivotal roles in cancer development and generated a mouse scFv (mscFv1C9) could effectively prohibit cancer cell proliferation in vitro and in vivo. Here, we further humanized this scFv (hscFv1C9) using a structure-guided complementarity determining region grafting strategy. The purified hscFv1C9 maintained similar antigen-binding affinity and specificity as mscFv1C9, and it was capable of inhibiting growth of different tumours in vitro and in vivo...
March 24, 2018: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/29563896/screening-of-a-scfv-antibody-with-high-affinity-for-application-in-human-ifn-%C3%AE-immunoassay
#18
Hang Yang, Yanfang Zhong, Juncheng Wang, Qinghong Zhang, Xiulan Li, Sumei Ling, Shihua Wang, Rongzhi Wang
Interferon gamma (IFN-γ), a signal proinflammatory cytokine secreted by immune cell, and plays a critical role in the pathogenesis and progression of many diseases. It has been regarded as an important marker for determination of disease-specific immune responses. Therefore, it is urgent to develop a feasible and accurate method to detect IFN-γ in clinic real blood samples. Until now, the immunoassay based on singe chain variable fragment (scFv) antibody for human IFN-γ is still not reported. In the present study, an scFv antibody named scFv-A8 with high specificity was obtained by phage display and biopanning, with the affinity 2...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29563774/preclinical-pharmacokinetics-interspecies-scaling-and-pharmacokinetics-of-a-phase-i-clinical-trial-of-ttac-0001-a-fully-human-monoclonal-antibody-against-vascular-endothelial-growth-factor-2
#19
Weon Sup Lee, Sang Ryeol Shim, Seon Young Lee, Jin San Yoo, Sung Kweon Cho
Background: VEGF is a highly selective mitogen that serves as the central regulator of tumor angiogenesis by mediating endothelial proliferation, permeability, and survival. Tanibirumab (TTAC-0001) is a fully human IgG1 monoclonal antibody derived from a fully human naïve single-chain variable fragment (ScFv) phage library that was developed to inhibit the effects of VEGF in the treatment of solid tumors, especially those of the brain. Methods: In the present study, we conducted intravenous pharmacokinetic studies of TTAC-0001 in mice, rats, and cynomolgus monkeys...
2018: Drug Design, Development and Therapy
https://www.readbyqxmd.com/read/29560124/cd30-targeted-oncolytic-viruses-as-novel-therapeutic-approach-against-classical-hodgkin-lymphoma
#20
Julia D S Hanauer, Benjamin Rengstl, Dina Kleinlützum, Johanna Reul, Anett Pfeiffer, Thorsten Friedel, Irene C Schneider, Sebastian Newrzela, Martin-Leo Hansmann, Christian J Buchholz, Alexander Muik
Classical Hodgkin lymphoma (cHL) is a hematopoietic malignancy with a characteristic cellular composition. The tumor mass is made up of infiltrated lymphocytes and other cells of hematologic origin but only very few neoplastic cells that are mainly identified by the diagnostic marker CD30. While most patients with early stage cHL can be cured by standard therapy, treatment options for relapsed or refractory cHL are still not sufficient, although immunotherapy-based approaches for the treatment of cHL patients have gained ground in the last decade...
February 27, 2018: Oncotarget
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