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https://www.readbyqxmd.com/read/27902325/targeting-naturally-occurring-epitope-variants-of-hepatitis-c-virus-with-high-affinity-t-cell-receptors
#1
Huajun Zhang, Jianbing Zhang, Lei Chen, Zhiming Weng, Ye Tian, Haifeng Zhao, Youjia Li, Lin Chen, Zhaoduan Liang, Hongjun Zheng, Wenzhuo Zhao, Shi Zhong, Yi Li
Hepatitis C virus (HCV) readily establishes chronic infection, which is characterized by failure of virus-specific CD8+ T cells. HCV uses epitope mutation and T-cell exhaustion to escape from the host immune response. Previously, we engineered high-affinity T-cell receptors (HATs) targeting HIV escape mutants. In this study, the affinity of a T-cell receptor specific for the human leukocyte antigen (HLA)-A2-restricted HCV immunodominant epitope NS3 1406-1415 (KLVALGINAV) was improved from a KD of 6.6 µM to 40 pM...
November 11, 2016: Journal of General Virology
https://www.readbyqxmd.com/read/27899028/nano-liposome-based-target-toxicity-machine-an-alternative-complementary-approach-in-atopic-diseases
#2
Behzad Mansoori, Ali Mohammadi, Neda Shajari, Sadaf Davudian, Shima Salehi, Behzad Baradaran
Despite using lots of progression in hypersensitivity, it is not yet a cure. Current treatment for allergy focuses on two approaches, including treats only the symptoms of allergy by utilizing drugs and the other is desensitization therapy (immunotherapy), which involves administration of specific allergens. The main obstacle to hypersensitivity permanent treatment is the IgE producing subclass of B-lymphocytes which is the source of IgE- producing in hypersensitive patients. Removal of these lymphocytes causes noticeable reduction in the levels of IgE and allergic responses significantly...
November 29, 2016: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/27894092/a-novel-fully-human-anti-ncl-immunornase-for-triple-negative-breast-cancer-therapy
#3
Chiara D'Avino, Dario Palmieri, Ashley Braddom, Nicola Zanesi, Cindy James, Sara Cole, Francesco Salvatore, Carlo M Croce, Claudia De Lorenzo
Breast cancer is the most common cancer in women worldwide. A new promising anti-cancer therapy involves the use of monoclonal antibodies specific for target tumor-associated antigens (TAAs). A TAA of interest for immunotherapy of Triple Negative Breast Cancer (TNBC) is nucleolin (NCL), a multifunctional protein, selectively expressed on the surface of cancer cells, which regulates the biogenesis of specific microRNAs (miRNAs) involved in tumor development and drug-resistance. We previously isolated a novel human anti-NCL scFv, called 4LB5, that is endowed with selective anti-tumor effects...
November 23, 2016: Oncotarget
https://www.readbyqxmd.com/read/27888365/development-of-a-novel-engineered-antibody-targeting-neisseria-species
#4
Chih-Yueh Liu, Chang-Ching Weng, Chih-Hsiang Lin, Chiou-Ying Yang, Kwok-Kong Tony Mong, Yaw-Kuen Li
OBJECTIVES: A Neissaria bacterial pilus sugar, bacillosamine, was synthesized and, for the first time, used as a probe to screen a single-chain variable fragment (scFv). RESULTS: Four Neisseria, Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria sicca and Neisseria subflava, and two negative controls, Streptococcus pneumoniae and Escherichia coli, were tested through ELISA, immunostaining and gold nanoparticle immunological assay. All results indicated that the selected scFv is feasible for the specific detection of Neisseria species via the recognition of bacillosamine...
November 25, 2016: Biotechnology Letters
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
#5
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...
November 22, 2016: Life Sciences
https://www.readbyqxmd.com/read/27881659/glycosyl-phosphatidylinositol-gpi-anchored-anti-hiv-scfv-efficiently-protect-cd4-t-cells-from-hiv-1-infection-and-deletion-in-hu-pbl-mice
#6
Chaobaihui Ye, Weiming Wang, Liang Cheng, Guangming Li, Michael Wen, Qi Wang, Qing Zhang, Dan Li, Paul Zhou, Lishan Su
: Despite the success in viral inhibition and CD4 T cell recovery by highly active antiretroviral treatment (HAART), HIV-1 is still not curable due to the persistence of HIV-1 reservoir during treatment. One patient with acute myeloid leukemia who received allogeneic hematopoietic stem cell transplantation from homozygous CCR5 Δ32 donor has had no detectable viremia for 9 years after HAART cessation. This case has inspired a field of HIV-1 cure research focusing on engineering HIV-1 resistance of permissive cells...
November 23, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27879646/integrating-scfv-into-xmap-assays-for-the-detection-of-marine-toxins
#7
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/27871972/recombinant-antibody-fragment-production
#8
REVIEW
H Ma, R O'Kennedy
Recombinant antibodies are now very important in both therapeutics and diagnostics and offer significant advantages over conventional antibodies. The generation of a single-chain variable antibody fragment (scFv) (a common and important recombinant antibody format) is used to demonstrate the construction of a recombinant antibody library. An immunotube-based two-day panning approach, using Escherichia coli as an expression system, is utilised for antibody screening. The methods used for antibody selection and purification using immobilized metal affinity chromatography (IMAC) are described...
November 18, 2016: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/27871874/functional-and-immuno-reactive-characterization-of-a-previously-undescribed-peptide-from-the-venom-of-the-scorpion-centruroides-limpidus
#9
Timoteo Olamendi-Portugal, Rita Restano-Cassulini, Lidia Riaño-Umbarila, Baltazar Becerril, Lourival D Possani
A previously undescribed toxic peptide named Cl13 was purified from the venom of the Mexican scorpion Centruroides limpidus. It contains 66 amino acid residues, including four disulfide bonds. The physiological effects assayed in 7 different subtypes of voltage gated Na(+)-channels, showed that it belongs to the β-scorpion toxin type. The most notorious effects were observed in subtypes Nav1.4, Nav1.5 and Nav1.6. Although having important sequence similarities with two other lethal toxins from this scorpion species (Cll1m and Cll2), the recently developed single chain antibody fragments (scFv) of human origin were not capable of protecting against Cl13...
November 18, 2016: Peptides
https://www.readbyqxmd.com/read/27861173/construction-expression-and-activity-of-a-novel-immunotoxin-comprising-a-humanized-antiepidermal-growth-factor-receptor-scfv-and-modified-pseudomonas-aeruginosa-exotoxin-a
#10
Bahman Akbari, Safar Farajnia, Nosratollah Zarghami, Nejat Mahdieh, Mohammad Rahmati, Shiva A Khosroshahi, Abolfazl Barzegar, Leila Rahbarnia
Overexpression of epidermal growth factor receptor (EGFR) plays a significant role in the development and metastasis of many solid tumors. Strategies based on anti-EGFR immunotoxins have shown promising results in several studies, but immunogenicity of antibody and toxin moieties is a limitation of this type of therapeutics. In the present study, a novel humanized anti-EGFR immunotoxin (huscFv-PE25) was developed by genetic fusing of a humanized anti-EGFR single-chain variable fragment (huscFv) with a modified Pseudomonas aeruginosa exotoxin A (PE25KDEL)...
November 16, 2016: Anti-cancer Drugs
https://www.readbyqxmd.com/read/27859059/influence-of-e-coli-chaperone-dnak-on-protein-immunogenicity
#11
Kirsty D Ratanji, Jeremy P Derrick, Ian Kimber, Robin Thorpe, Meenu Wadhwa, Rebecca J Dearman
The production of anti-drug antibodies can impact significantly upon the safety and efficacy of biotherapeutics. It is known that various factors, including aggregation and the presence of process-related impurities can modify and augment the immunogenic potential of proteins. The purpose of the investigations reported here was to characterize in mice the influence of aggregation and host cell protein (HCP) impurities on the immunogenicity of a humanized single chain antibody variable fragment (scFv), and mouse albumin...
November 10, 2016: Immunology
https://www.readbyqxmd.com/read/27856232/improved-recombinant-antibody-production-by-cho-cells-using-a-production-enhancer-dna-element-with-repeated-transgene-integration-at-a-predetermined-chromosomal-site
#12
Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Takeshi Omasa, Masamichi Kamihira
Chinese hamster ovary (CHO) cells are one of the most useful host cell lines for the production of biopharmaceutical proteins. Although a series of production processes have been refined to improve protein productivity and cost performance, establishing producer cells is still time-consuming and labor-intensive. Recombinase-mediated site-specific gene integration into a predetermined chromosomal locus may enable predictable protein expression, reducing the laborious process of cell screening. We previously developed an accumulative site-specific gene integration system (AGIS) using Cre recombinase and mutated loxP sites for transgene integration and amplification in the CHO cell genome...
November 14, 2016: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/27854147/improving-the-developability-of-an-anti-epha2-single-chain-variable-fragment-for-nanoparticle-targeting
#13
Melissa L Geddie, Neeraj Kohli, Dmitri B Kirpotin, Maja Razlog, Yang Jiao, Tad Kornaga, Rachel Rennard, Lihui Xu, Birgit Schoerberl, James D Marks, Daryl C Drummond, Alexey A Lugovskoy
Antibody-targeted nanoparticles have great promise as anti-cancer drugs; however, substantial developmental challenges of antibody modules prevent many candidates from reaching the clinic. Here, we describe a robust strategy for developing an EphA2-targeting antibody fragment for immunoliposomal drug delivery. A highly bioactive single-chain variable fragment (scFv) was engineered to overcome developmental liabilities, including low thermostability and weak binding to affinity purification resins. Improved thermostability was achieved by modifying the framework of the scFv, and complementarity-determining region (CDR)-H2 was modified to increase binding to protein A resins...
November 17, 2016: MAbs
https://www.readbyqxmd.com/read/27853644/vectorization-in-an-oncolytic-vaccinia-virus-of-an-antibody-a-fab-and-a-scfv-against-programmed-cell-death-1-pd-1-allows-their-intratumoral-delivery-and-an-improved-tumor-growth-inhibition
#14
Patricia Kleinpeter, Laetitia Fend, Christine Thioudellet, Michel Geist, Nathalie Sfrontato, Véronique Koerper, Catherine Fahrner, Doris Schmitt, Murielle Gantzer, Christelle Remy-Ziller, Renée Brandely, Dominique Villeval, Karola Rittner, Nathalie Silvestre, Philippe Erbs, Laurence Zitvogel, Eric Quéméneur, Xavier Préville, Jean-Baptiste Marchand
We report here the successful vectorization of a hamster monoclonal IgG (namely J43) recognizing the murine Programmed cell death-1 (mPD-1) in Western Reserve (WR) oncolytic vaccinia virus. Three forms of mPD-1 binders have been inserted into the virus: whole antibody (mAb), Fragment antigen-binding (Fab) or single-chain variable fragment (scFv). MAb, Fab and scFv were produced and assembled with the expected patterns in supernatants of cells infected by the recombinant viruses. The three purified mPD-1 binders were able to block the binding of mPD-1 ligand to mPD-1 in vitro...
2016: Oncoimmunology
https://www.readbyqxmd.com/read/27849134/developing-and-characterization-of-single-chain-variable-fragment-scfv-antibody-against-frizzled-7-fzd7-receptor
#15
Hamid Nickho, Vahid Younesi, Leili Aghebati-Maleki, Morteza Motallebnezhad, Jafar Majidi Zolbanin, Aliakbar Movassagh Pour, Mehdi Yousefi
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/27845043/molecular-characterization-of-a-single-chain-antibody-variable-fragment-scfv-specific-for-pspa-from-streptococcus-pneumoniae
#16
ShinA Jang, GyuHee Kim, Jee Hye Oh, Seung Yeob Rhee, Dongho Kim, Kook Han Kim, Yong Ho Kim, Dong-Kwon Rhee, Sangho Lee
Streptococcus pneumoniae is a major infectious agent responsible for pneumonia, otitis media, sepsis and meningitis. Pneumococcal surface protein A (PspA) is a well-characterized virulence factor localized on the surface and a target for vaccine development. In this study, we screened a single-chain antibody variable fragment (scFv) using phage display from a human synthetic library to select a clone 2B11. Affinity (Kd) of 2B11 was measured to be 5 nM using biolayer interferometry. 2B11 exhibited a dose-dependent recognition of recombinant PspA with no cross-reactivity towards pneumococcal antigens...
November 11, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27836986/targeting-thrombomodulin-to-circulating-red-blood-cells-augments-its-protective-effects-in-models-of-endotoxemia-and-ischemia-reperfusion-injury
#17
Ronald Carnemolla, Carlos H Villa, Colin F Greineder, Sergei Zaitsev, Kruti R Patel, M Anna Kowalska, Dmitriy N Atochin, Douglas B Cines, Don L Siegel, Charles T Esmon, Vladimir R Muzykantov
Endothelial thrombomodulin (TM) regulates coagulation and inflammation via several mechanisms, including production of activated protein C (APC). Recombinant APC and soluble fragments of TM (sTM) have been tested in settings associated with insufficiency of the endogenous TM/APC pathway, such as sepsis. We previously designed a fusion protein of TM [single-chain variable fragment antibody (scFv)/TM] targeted to red blood cells (RBCs) to improve pharmacokinetics and antithrombotic effects without increasing bleeding...
November 11, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/27834948/the-antitumor-efficacy-of-anti-p21ras-scfv-mediated-by-the-dual-promoter-regulated-recombinant-adenovirus-kghv300
#18
X Pan, X Liu, J Li, S Zhen, D Liu, Q Feng, W Zhao, Y Luo, Y Zhang, H Li, J Yang
Ras mutations and overexpression of the Ras protein; i.e., p21Ras, are main causes of cancer development and progression, which has made the Ras gene and p21Ras important targets for therapy of Ras-driven cancers. We previously prepared recombinant adenovirus KGHV100 based on replication-defective adenovirus type 5, which could intracellularly express anti-p21Ras single chain fragment viable antibodies (scFv) and repress tumor growth in vitro and in vivo. However, the anti-tumor effects of this anti-p21Ras scFv were limited by short-term scFv expression due to a replication defect of KGHV100...
November 11, 2016: Gene Therapy
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
#19
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/27825868/development-of-a-targeted-anti-her2-scfv-chimeric-peptide-for-gene-delivery-into-her2-positive-breast-cancer-cells
#20
Roya Cheraghi, Mahboobeh Nazari, Mohsen Alipour, Asia Majidi, Saman Hosseinkhani
Chimeric polymers are known as suitable carriers for gene delivery. Certain properties are critical for a polymer to be used as a gene delivery vector. A new polymer was designed for the targeted delivery of genes into breast cancer cell lines, based on MPG peptide. It is composed of different functional domains, including HIV gp41, nuclear localization sequence of SV40 T-antigen, two C-terminus repeats of histone H1, and the scFv of anti-HER2 antibody. The results demonstrated that the vector can effectively condense plasmid DNA into nanoparticles with an average size of 250nm...
November 5, 2016: International Journal of Pharmaceutics
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