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scfv phage display

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https://www.readbyqxmd.com/read/28101463/development-of-a-novel-human-single-chain-antibody-against-egfrviii-antigen-by-phage-display-technology
#1
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Bahman Akbari, Shiva Ahdi Khosroshahi
Purpose: EGFRvIII as the most common mutant variant of the epidermal growth factor receptor is resulting from deletion of exons 2-7 in the coding sequence and junction of exons 1 and 8 through a novel glycine residue. EGFRvIII is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. The aim of the present study was identification of a novel single chain antibody against EGFRvIII as a promising target for cancer therapy. Methods: In this study, a synthetic peptide corresponding to EGFRvIII protein was used for screening a naive human scFv phage library...
December 2016: Advanced Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28095447/identification-and-characterization-of-single-chain-antibodies-that-specifically-bind-gi-noroviruses
#2
Amy M Hurwitz, Wanzhi Huang, Baijun Kou, Mary K Estes, Robert L Atmar, Timothy Palzkill
Norovirus infections commonly lead to outbreaks of acute gastroenteritis and spread quickly, resulting in many health and economic challenges prior to diagnosis. Rapid and reliable diagnostic tests are therefore essential to identify infections and to guide the appropriate clinical responses at the point-of-care. Existing tools, including RT-PCR and enzyme immunoassays, pose several limitations based on the significant time, equipment and expertise required to elicit results. Immunochromatographic assays available for use at the point-of-care have poor sensitivity and specificity, especially for genogroup I noroviruses, thus requiring confirmation of results with more sensitive testing methods...
2017: PloS One
https://www.readbyqxmd.com/read/28090795/isolation-and-characterization-of-anti-c-met-single-chain-fragment-variable-scfv-antibodies
#3
Elmira Safaie Qamsari, Zahra Sharifzadeh, Salman Bagheri, Farhad Riazi-Rad, Vahid Younesi, Mohsen Abolhassani, Sepideh Safaei Ghaderi, Behzad Baradaran, Mohammad Hossein Somi, Mehdi Yousefi
The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques...
January 16, 2017: Journal of Immunotoxicology
https://www.readbyqxmd.com/read/28075174/analysis-of-sivmac-envelope-specific-antibodies-selected-via-phage-display
#4
Sergio Ita, Mayara Rovariz Agostinho, Katherine Sullivan, Seung Yub Han, Rana Akleh, Welkin Johnson, Ismael Ben Farouck Fofana
We have constructed a single chain Fv (scFv) phage display library from an SIV-infected rhesus macaque that developed unusually high-titer neutralizing antibody responses against tier-3, neutralization-resistant SIVmac239. The library was screened using trimeric (gp140) and monomeric (gp120) forms of the SIVmac239 envelope (Env) glycoprotein. We also cloned variable-heavy and variable-light (VH-VL) antibody fragments from 7 previously described rhesus macaque B-cell lines (BLCL) that produce SIV gp120-specific monoclonal antibodies (mAbs)...
January 11, 2017: AIDS Research and Human Retroviruses
https://www.readbyqxmd.com/read/28075071/multi-target-selection-of-catalytic-antibodies-wih-%C3%AE-lactamase-activity-using-phage-display
#5
Melody A Shahsavarian, Nancy Chaaya, Narciso Costa, Didier Boquet, Alexandre Atkinson, Bernard Offmann, Srini V Kaveri, Sébastien Lacroix-Desmazes, Alain Friboulet, Bérangère Avalle, Séverine Padiolleau-Lefèvre
β-lactamase enzymes responsible for bacterial resistance to antibiotics are among the most important health threats to the human population today. Understanding the increasingly vast structural motifs responsible for the catalytic mechanism of β-lactamases will help improve the future design of new generation antibiotics and mechanism-based inhibitors of these enzymes. Here we report the construction of a large murine scFv phage display library of size 2.7×10(9) with extended diversity by combining different mouse models...
January 11, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28064157/fviii-specific-human-chimeric-antigen-receptor-car-t-regulatory-cells-suppress-t-and-b-cell-responses-to-fviii
#6
Jeongheon Yoon, Anja Schmidt, Ai-Hong Zhang, Christoph Königs, Yong Chan Kim, David W Scott
Replacement therapy with factor VIII (FVIII) is used in hemophilia A patients for treatment of bleeding episodes or for prophylaxis. A common and serious problem with this therapy is the patient's immune response to FVIII, due to a lack of tolerance, leading to the formation of inhibitory antibodies. Development of tolerogenic therapies, other than standard ITI, is an unmet goal. We previously generated engineered antigen- specific regulatory T cells (Tregs), created by transduction of a recombinant T cell receptor (TCR) isolated from a hemophilia A subject's T cell clone...
November 15, 2016: Blood
https://www.readbyqxmd.com/read/27984065/isolation-and-characterization-of-a-novel-human-scfv-inhibiting-egfr-viii-expressing-cancers
#7
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Hassan Dariushnejad, Mohammad Kazem Hosseini
EGFRvIII, a mutant form of epidermal growth factor receptor is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. This tumor specific antigen has emerged as a promising candidate for antibody based therapy of several cancers. The aim of the present study was isolation and characterization of a human single chain antibody against EGFRvIII as a promising target for cancer therapy. For this, a synthetic peptide corresponding to EGFRvIII protein was used for screening the naive human scFv phage library...
October 29, 2016: Immunology Letters
https://www.readbyqxmd.com/read/27956240/array-in-well-binding-assay-for-multiparameter-screening-of-phage-displayed-antibodies
#8
Susan Pérez-Gamarra, Liisa Hattara, Gaurav Batra, Petri Saviranta, Urpo Lamminmäki
Phage display is a well-established and powerful tool for the development of recombinant antibodies. In a standard phage display selection process using a high quality antibody phage library, a large number of unique antibody clones can be generated in short time. However, the pace of the antibody discovery project eventually depends on the methodologies used in the next screening phase to identify the clones with the most promising binding characteristics e.g., in terms of specificity, affinity and epitope...
December 9, 2016: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/27942613/characterization-of-mab6-9-1-monoclonal-antibody-against-hemagglutinin-of-avian-influenza-virus-h5n1-and-its-engineered-derivative-single-chain-variable-fragment-antibody
#9
Róża Sawicka, Paweł Siedlecki, Barbara Kalenik, Jan P Radomski, Violetta Sączyńska, Anna Porębska, Bogusław Szewczyk, Agnieszka Sirko, Anna Góra-Sochacka
Hemagglutinin (HA), as a major surface antigen of influenza virus, is widely used as a target for production of neutralizing antibodies. Monoclonal antibody, mAb6-9-1, directed against HA of highly pathogenic avian influenza virus A/swan/Poland/305-135V08/2006(H5N1) was purified from mouse hybridoma cells culture and characterized. The antigenic specificity of mAb6-9-1 was verified by testing its cross-reactivity with several variants of HA. The mimotopes recognized by mAb6-9-1 were selected from two types of phage display peptide libraries...
December 7, 2016: Acta Biochimica Polonica
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
#10
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/27849134/developing-and-characterization-of-single-chain-variable-fragment-scfv-antibody-against-frizzled-7-fzd7-receptor
#11
Hamid Nickho, Vahid Younesi, Leili Aghebati-Maleki, Morteza Motallebnezhad, Jafar Majidi Zolbanin, Aliakbar Movassagh Pour, Mehdi Yousefi
ABSTACT 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
#12
ShinA Jang, Gyuhee Kim, Jihye Oh, Seungyeop Lee, 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...
January 1, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27792006/construction-of-human-anti-tetanus-single-chain-variable-fragment-applying-symplex-technology
#13
Laya Esmati, Jalil Fallah Mehrabadi, Masoumeh Bazaz, Hamideh Rouhani Nejad
BACKGROUND: Human monoclonal antibodies are important molecules in clinical research. Current Limitations of mAb technologies namely instability of immortalized B-cell line and probability of forming unusual VH-VL pairs in phage-display method led to mAbs technology based on single plasma cell called ``SYMPLEX''. OBJECTIVE: In this method, cognate VH and VL fragments generated from individual antibody genes exactly the same as natural ones. METHODS: PBMCs of whole blood of an immunized candidate was used as a resource of rearranged Ab genes...
October 20, 2016: Human Antibodies
https://www.readbyqxmd.com/read/27786270/recombinant-antibodies-for-specific-detection-of-clostridial-fe-fe-hydrogenases
#14
Rahul Mangayil, Matti Karp, Urpo Lamminmäki, Ville Santala
Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels...
October 27, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27779085/development-of-a-novel-human-scfv-against-egfr-l2-domain-by-phage-display-technology
#15
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Kamal Veisi, Shiva Ahdi Khosroshahi, Asghar Tanomand
Epidermal growth factor receptor (EGFR) as a transmembrane tyrosine kinase receptor is frequently overexpresses in tumors with epithelial origin. The L2 domain from extracellular part of EGFR is involved in ligand binding and the blockage of this domain prevents activation of related signaling pathways. This study was aimed to develop a novel human scFv against EGFR L2 domain as a promising target for cancer therapy. The L2 recombinant protein was purified and used for panning a human scFv phage library (Tomlinson I)...
September 28, 2016: Current Pharmaceutical Design
https://www.readbyqxmd.com/read/27730560/optimized-generation-of-high-affinity-high-specificity-single-chain-fv-antibodies-from-multi-antigen-immunized-chickens
#16
William J J Finlay, Laird Bloom, Sreeja Varghese, Bénédicte Autin, Orla Cunningham
High-affinity, highly specific binding proteins are a key class of molecules used in the development of new affinity chromatography methods. Traditionally, antibody-based methods have relied on the use of immunoglobulins purified from immune animal sera, from egg yolks, or from murine monoclonal hybridoma supernatants. To accelerate and refine the reagent antibody generation process, we have developed optimized methods that allow the rapid assembly of scFv libraries from chickens immunized with pools of immunogens...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27663029/production-and-characterization-of-neutralizing-antibodies-against-bungarus-multicinctus-snake-venom
#17
Chi-Hsin Lee, Yu-Ching Lee, Sy-Jye Leu, Liang-Tzung Lin, Jen-Ron Chiang, Wei-Jane Hsu, Yi-Yuan Yang
: Banded krait (Bungarus multicinctus; BM), one of the major envenomation species in Taiwan, contains neurotoxic venom proteins (BM proteins), that poses a serious medical problem in tropical and sub-tropical countries. Even though horse-derived serum is an efficient therapy against snake venom, it is associated with high cost and side effects. Therefore, developing a more cost-effective alternative treatment option is highly envisaged. In this study, chickens were immunized with BM proteins, and polyclonal immunoglobulin Y (IgY) antibodies were purified from eggs...
September 23, 2016: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/27626445/tetanus-neurotoxin-neutralizing-antibodies-screened-from-a-human-immune-scfv-antibody-phage-display-library
#18
Han Wang, Rui Yu, Ting Fang, Ting Yu, Xiangyang Chi, Xiaopeng Zhang, Shuling Liu, Ling Fu, Changming Yu, Wei Chen
Tetanus neurotoxin (TeNT) produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc) as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H) can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro...
2016: Toxins
https://www.readbyqxmd.com/read/27622814/screening-and-molecular-evolution-of-a-single-chain-variable-fragment-antibody-scfv-against-citreoviridin-toxin
#19
Rongzhi Wang, Xiaosong Gu, Zhenghong Zhuang, Yanfang Zhong, Hang Yang, Shihua Wang
Citreoviridin (CIT), a small food-borne mycotoxin produced by Penicillium citreonigrum, is generally distributed in various cereal grains and farm crop products around the world and has caused cytotoxicity as an uncompetitive inhibitor of ATP hydrolysis. A high affinity single chain variable fragment (scFv) antibody that can detect the citreoviridin in samples is still not available; therefore, it is very urgent to prepare an antibody for CIT detection and therapy. In this study, an amplified and assembled scFv from hybridoma was used to construct the mutant phage library by error-prone PCR, generating a 2 × 10(8) capacity mutated phage display library...
October 12, 2016: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/27590051/generation-and-analyses-of-human-synthetic-antibody-libraries-and-their-application-for-protein-microarrays
#20
Anna Säll, Maria Walle, Christer Wingren, Susanne Müller, Tomas Nyman, Andrea Vala, Mats Ohlin, Carl A K Borrebaeck, Helena Persson
Antibody-based proteomics offers distinct advantages in the analysis of complex samples for discovery and validation of biomarkers associated with disease. However, its large-scale implementation requires tools and technologies that allow development of suitable antibody or antibody fragments in a high-throughput manner. To address this we designed and constructed two human synthetic antibody fragment (scFv) libraries denoted HelL-11 and HelL-13. By the use of phage display technology, in total 466 unique scFv antibodies specific for 114 different antigens were generated...
October 2016: Protein Engineering, Design & Selection: PEDS
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