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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
#1
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
#2
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
#3
ShinA Jang, Gyuhee Kim, Jihye Oh, Seungyeob 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...
November 11, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27792006/construction-of-human-anti-tetanus-single-chain-variable-fragment-applying-symplex-technology
#4
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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
#11
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
https://www.readbyqxmd.com/read/27581963/screening-of-functional-antibodies-by-a-scfv-phage-display-library-system-for-innovative-biodrug-development
#12
Shin-Ichi Tsunoda
No abstract text is available yet for this article.
September 2016: Nihon Yakurigaku Zasshi. Folia Pharmacologica Japonica
https://www.readbyqxmd.com/read/27578501/continuous-microfluidic-assortment-of-interactive-ligands-cmail
#13
Yi-Hsing Hsiao, Chao-Yang Huang, Chih-Yung Hu, Yen-Yu Wu, Chung-Hsiun Wu, Chia-Hsien Hsu, Chihchen Chen
Finding an interactive ligand-receptor pair is crucial to many applications, including the development of monoclonal antibodies. Biopanning, a commonly used technique for affinity screening, involves a series of washing steps and is lengthy and tedious. Here we present an approach termed continuous microfluidic assortment of interactive ligands, or CMAIL, for the screening and sorting of antigen-binding single-chain variable antibody fragments (scFv) displayed on bacteriophages (phages). Phages carrying native negative charges on their coat proteins were electrophoresed through a hydrogel matrix functionalized with target antigens under two alternating orthogonal electric fields...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27558409/novel-nanoscale-bacteriophage-based-single-domain-antibodies-for-the-therapy-of-systemic-infection-caused-by-candida-albicans
#14
Shuai Dong, Hongxi Shi, Donghui Cao, Yicun Wang, Xintong Zhang, Yan Li, Xiang Gao, Li Wang
Candida albicans (C. albicans) is an important human commensal and opportunistic fungal pathogen. Secreted aspartyl proteinases (Saps) are a major virulence trait of C. albicans, and among these proteases Sap2 has the highest expression levels. It is possible that antibodies against Sap2 could provide an antifungal effect. In this study, two phages displaying anti-rSap2 single chain variable fragments (scFvs) were screened from human single fold scFv libraries, and their potential therapeutic roles were evaluated using a murine model infected by C...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27550798/high-throughput-cytotoxicity-screening-of-anti-her2-immunotoxins-conjugated-with-antibody-fragments-from-phage-displayed-synthetic-antibody-libraries
#15
Shin-Chen Hou, Hong-Sen Chen, Hung-Wei Lin, Wei-Ting Chao, Yao-Sheng Chen, Chi-Yu Fu, Chung-Ming Yu, Kai-Fa Huang, Andrew H-J Wang, An-Suei Yang
Immunotoxins are an important class of antibody-based therapeutics. The potency of the immunotoxins depends on the antibody fragments as the guiding modules targeting designated molecules on cell surfaces. Phage-displayed synthetic antibody scFv libraries provide abundant antibody fragment candidates as targeting modules for the immunoconjugates, but the discovery of optimally functional immunoconjugates is limited by the scFv-payload conjugation procedure. In this work, cytotoxicity screening of non-covalently assembled immunotoxins was developed in high throughput format to discover highly functional synthetic antibody fragments for delivering toxin payloads...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27520285/invert-biopanning-a-novel-method-for-efficient-and-rapid-isolation-of-scfvs-by-phage-display-technology
#16
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Kamal Veisi, Asghar Tanomand, Bahman Akbari
Phage display is a prominent screening technique for development of novel high affinity antibodies against almost any antigen. However, removing false positive clones in screening process remains a challenge. The aim of this study was to develop an efficient and rapid method for isolation of high affinity scFvs by removing NSBs without losing rare specific clones. Therefore, a novel two rounds strategy called invert biopanning was developed for isolating high affinity scFvs against EGFRvIII antigen from human scFv library...
August 9, 2016: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/27472865/design-and-construction-of-immune-phage-antibody-library-against-tetanus-neurotoxin-production-of-single-chain-antibody-fragments
#17
Sanam Sadreddini, Mehrnosh Seifi-Najmi, Babollah Ghasemi, Hossein Samadi Kafil, Vahideh Alinejad, Sevil Sadreddini, Vahid Younesi, Farhad Jadidi-Niaragh, Mehdi Yousefi
BACKGROUND: Tetanus neurotoxin (TeNT) is composed of a light (LC) and heavy chain (HC) polypeptides, released by anaerobic bacterium Clostridium tetani and can cause fatal life-threatening infectious disease. Toxin HC and LC modules represents receptor binding and zinc metalloprotease activity, respectively. The passive administration of animal-derived antibodies against tetanus toxin has been considered as the mainstay therapy for years. However, this treatment is associated with several adverse effects due to the presence of anti-isotype antibodies...
December 23, 2015: Human Antibodies
https://www.readbyqxmd.com/read/27450754/next-generation-sequencing-of-all-variable-loops-of-synthetic-single-framework-scfv-application-in-anti-hdl-antibody-selections
#18
Janita Lövgren, Juha-Pekka Pursiheimo, Mikko Pyykkö, Jussi Salmi, Urpo Lamminmäki
Next generation sequencing (NGS) can be applied to monitoring antibody phage display library selection processes to follow the enrichment of each individual antibody clone. Utilising the recent development of the Illumina sequencing platform enabling sequencing up to 2×300bp, we have developed a method to deep sequence all complementarity determining regions (CDRs) in the clones obtained from a synthetic single framework antibody library. This was complemented by an in-house bioinformatics pipeline for efficient analysis of the sequencing results...
December 25, 2016: New Biotechnology
https://www.readbyqxmd.com/read/27411546/development-of-a-biotinylated-broad-specificity-single-chain-variable-fragment-antibody-and-a-sensitive-immunoassay-for-detection-of-organophosphorus-pesticides
#19
Fengchun Zhao, Yuan Tian, Huimin Wang, Jiye Liu, Xiao Han, Zhengyou Yang
Organophosphorus pesticides (OPs) are the most widely used pesticides in agriculture, and OP residues have been broadly reported in food and environmental samples. The aim of this study is to develop a recombinant antibody-based broad-specificity immunoassay for OPs. A phage display library was prepared from a mouse pre-immunized with a generic immunogen of OPs, and a single-chain variable fragment (scFv) antibody was selected. The selected scFv antibody was fused with biotin acceptor domain (BAD) and overexpressed as an inclusion body in Escherichia coli BL21 (DE3)...
September 2016: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/27371853/-construction-and-panning-of-scfv-phage-display-library-against-recombinant-interleukin-4-receptor
#20
Guangyong Yang, Haitao Guo, Ximing Liu, Guangzhi He, Weiyi Tian, Kun Cai, Ping Wang, Wenjia Wang
Objective To construct the recombinant human interleukin 4 receptor (rhIL-4R) single-chain Fv (scFv) antibody library by phage display technique to obtain the anti-IL-4R scFv clones selected from the library. Methods Total RNA was extracted from splenocytes of the BALB/c mice immunized with rhIL-4R. Complementary DNA fragments of variable heavy (VH) and variable light (VL) chains of the antibodies were prepared by reverse transcription PCR and assembled into scFv by splice overlap extension PCR (SOE-PCR). Both scFv and the pCANTAB5E vector were respectively double-digested with restriction endonuclease Sfi I and Not I, connected with T4 ligase, and then transformed into the competent cells E...
June 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
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