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scfv phage display

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https://www.readbyqxmd.com/read/29351623/development-of-sugar-chain-binding-single-chain-variable-fragment-antibody-to-adult-t-cell-leukemia-cells-using-glyco-nanotechnology-and-phage-display-method
#1
Kaname Muchima, Taro Todaka, Hiroyuki Shinchi, Ayaka Sato, Arisa Tazoe, Rikiya Aramaki, Yuhei Kakitsubata, Risa Yokoyama, Naomichi Arima, Masanori Baba, Masahiro Wakao, Yuji Ito, Yasuo Suda
Adult T-cell leukemia (ATL) is an intractable blood cancer caused by the infection of human T-cell leukemia virus type-1, and effective medical treatment is required. It is known that the structure and expression levels of cell surface sugar chains vary depending on cell states such as inflammation and cancer. Thus, it is expected that the antibody specific for ATL cell surface sugar chain would be an effective diagnostic tool and a strong candidate for the development of an anti-ATL drug. Here, we developed a stable sugar chain binding single chain variable fragment antibody (scFv) that can bind to ATL cells using a fiber type Sugar Chip and phage display method...
January 17, 2018: Journal of Biochemistry
https://www.readbyqxmd.com/read/29330770/decoding-selection-bias-imparted-by-unpaired-cysteines-a-tug-of-war-between-expression-and-affinity
#2
B Vijayalakshmi Ayyar, Stephen Hearty, Richard O'Kennedy
In a recombinant antibody scFv format, the presence of an unpaired cysteine (Cys) is implicated in reduced soluble expression and inefficient presentation in phage display. Compared to other species, antibodies derived from rabbits are more likely to contain this unpaired Cys residue at position 80 (Cys80), when generated in a scFv format. In a screening campaign to isolate rabbit scFv against cardiac troponin I (cTnI), it was found that, a large proportion of isolated cTnI-specific clones contained unpaired Cys80...
January 12, 2018: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29315611/isolation-of-a-high-affinity-bet-v-1-specific-igg-derived-scfv-from-a-subject-vaccinated-with-hypoallergenic-bet-v-1-fragments
#3
Elisabeth Gadermaier, Katharina Marth, Christian Lupinek, Raffaela Campana, Gerhard Hofer, Katharina Blatt, Dubravka Smiljkovic, Uwe Roder, Margarete Focke-Tejkl, Susanne Vrtala, Walter Keller, Peter Valent, Rudolf Valenta, Sabine Flicker
BACKGROUND: Recombinant hypoallergenic allergen derivatives have been used in clinical immunotherapy studies and clinical efficacy seems to be related to the induction of blocking IgG antibodies recognizing the wild type allergens. However, so far no treatment-induced IgG antibodies have been characterized. OBJECTIVE: To clone, express and characterize IgG antibodies induced by vaccination with two hypoallergenic recombinant fragments of the major birch pollen allergen, Bet v 1 in a non-allergic subject...
January 9, 2018: Allergy
https://www.readbyqxmd.com/read/29306173/blocking-osteopontin-fibronectin-interactions-reduce-extracellular-fibronectin-deployment-and-arthritic-immunopathology
#4
Brij Bhushan Mehta, Saniya Sharma, Rakesh K Vasishta, Ramesh K Sen, Aman Sharma, Manni Luthra-Guptasarma
Elevated levels of a thrombin-cleaved fragment of osteopontin (OPNT) are seen in synovial fluid (SF) and tissues of rheumatoid arthritis (RA) patients. OPNT binds to integrins on cell surfaces, inducing adhesion, migration and survival of inflammatory cells in the synovial joints, where OPNT binds to fibronectin to link fibroblast-like synoviocytes (FLS) with B cells, stimulating the latter to produce inflammatory cytokines. Our aim was to block OPNT-fibronectin interactions and examine whether this reduces inflammation...
January 3, 2018: International Immunopharmacology
https://www.readbyqxmd.com/read/29305748/production-of-human-single-chain-fragment-antibody-scfv-against-human-epidermal-growth-factor-receptor-2-her-2-by-phage-display-technology
#5
Saeideh Foroumadi, Masoumeh Rajabibazl, Azam Rahimpour, Solmaz Shahidi, Walead Ebrahimizadeh, Maral Yarahmadi, Shirin Rajabi, Azam Daraei
Breast cancer with more than 1.7 million diagnoses per year has been known as one of the most prevalent cancers among women worldwide. Despite the availability of advanced treatment options, cancer progression and metastasis is observed in 20% of patients. Human epidermal growth factor receptor-2 (HER-2) is considered as an important prognostic and diagnostic tumor marker for breast cancer. While HER-2 is expressed on the surface of normal cells, its overexpression occurs in 20-25% on breast cancer tumor cells...
January 5, 2018: In Vitro Cellular & Developmental Biology. Animal
https://www.readbyqxmd.com/read/29199242/a-single-step-breeding-generated-a-diagnostic-anti-cortisol-antibody-fragment-with-over-30-fold-enhanced-affinity
#6
Hiroyuki Oyama, Izumi Morita, Yuki Kiguchi, Tomomi Morishita, Sakiko Fukushima, Yuki Nishimori, Toshifumi Niwa, Norihiro Kobayashi
Cortisol levels in bodily fluids represent a useful index for pituitary-adrenal function, and thus practical anti-cortisol antibodies are required. We have studied "antibody-breeding" approaches, which involve in vitro evolution of antibodies to improve their antigen-binding performances. Here, we produced an antibody fragment to measure serum cortisol levels with over 30-fold enhanced affinity after single mutagenesis and selection steps. A mouse anti-cortisol antibody, Ab-CS#3, with insufficient affinity for practical use, was chosen as the prototype antibody...
2017: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/29165857/isolation-of-phage-display-library-derived-scfv-antibody-specific-to-listeria-monocytogenes-by-a-novel-immobilized-method
#7
Nguyen Xuan-Hung, Thu-Le Trinh, Huong Thi-Bich Vu, Huan Quang Le, Anh To Kim
AIMS: To select Listeria monocytogenes specific scFv antibodies from a phage display library by a novel simple and cost-effective immobilization method. METHODS AND RESULTS: Light Expanded Clay Aggregate (LECA) was used as biomass support matrix for biopanning of a phage display library to select Listeria monocytogenes specific scFv antibody. Four rounds of positive selection against LECA-immobilized L. monocytogenes and an additional subtractive panning against L...
November 22, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29165352/antibody-based-protective-immunity-against-helminth-infections-antibody-phage-display-derived-antibodies-against-bmr1-antigen
#8
Anizah Rahumatullah, Izzati Zahidah Abdul Karim, Rahmah Noordin, Theam Soon Lim
Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and naïve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BmR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BmR1 antigen in healthy and infected states...
November 22, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29130184/generation-of-high-sensitivity-monoclonal-antibodies-specific-for-homoserine-lactones
#9
Soumya Palliyil
A number of bacteria use a class of chemical compounds called acyl-homoserine lactones (AHLs) as quorum sensing (QS) signals to coordinate their behavior at the population level, including pathogens like Pseudomonas aeruginosa. Blocking QS using antibodies is an attractive strategy for infection control as this process takes a central role in P. aeruginosa infections. Here the methods involved in the generation of high sensitivity anti-QS monoclonal antibodies from an immunized sheep phage display antibody library are described...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29119245/development-and-characterisation-of-a-novel-glucagon-like-peptide-1-receptor-antibody
#10
Emma K Biggs, Lihuan Liang, Jacqueline Naylor, Shimona Madalli, Rachel Collier, Matthew P Coghlan, David J Baker, David C Hornigold, Peter Ravn, Frank Reimann, Fiona M Gribble
AIMS/HYPOTHESIS: Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of GLP-1 remain unexplained at a mechanistic level, particularly in extrapancreatic tissues. The aim of this study was to generate and characterise a monoclonal antagonistic antibody for the GLP1R for use in vivo...
November 9, 2017: Diabetologia
https://www.readbyqxmd.com/read/29116510/parallelized-antibody-selection-in-microtiter-plates
#11
Giulio Russo, Doris Meier, Saskia Helmsing, Esther Wenzel, Fabian Oberle, André Frenzel, Michael Hust
The most common in vitro technology to generate human antibodies is phage display. This technology is a key technology to select recombinant antibodies for the use as research tools, in diagnostic tests, and for the development of therapeutics.In this review, the high-throughput compatible selection of antibodies (scFv) in microtiter plates is described. The given detailed protocols allow the antibody selection ("panning"), screening and identification of monoclonal antibodies in less than 1 week.
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29116508/construction-of-a-synthetic-antibody-gene-library-for-the-selection-of-intrabodies-and-antibodies
#12
Déborah Caucheteur, Gautier Robin, Vincent Parez, Pierre Martineau
Libraries of antibody fragments displayed on filamentous phages have proved their value to generate human antibodies against virtually any target. We describe here a simple protocol to make large and diverse libraries based on a single or a limited number of frameworks. The approach is flexible enough to be used with any antibody format, either single-chain (scFv, VHH) or multi-chain (Fv, Fab, (Fab')2), and to target in a single step the six complementarity-determining regions-or any other part-of the antibody molecule...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29116502/construction-of-bovine-immunoglobulin-libraries-in-the-single-chain-fragment-variable-scfv-format
#13
Ulrike S Diesterbeck
Recombinant immunoglobulins are an excellent tool for diagnosis, treatment, and passive immunization. Phage display offers a robust technique for the selection of recombinant antibodies from immunoglobulin libraries. The construction of immunoglobulin libraries for veterinary purposes was restricted by the lack of knowledge about species-specific diversities.The now available data enable the construction of highly diverse libraries in livestock like cattle. Using diverse primer sets, the immunoglobulin repertoire is amplified and ligated into a phagemid...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29116500/modular-construction-of-large-non-immune-human-antibody-phage-display-libraries-from-variable-heavy-and-light-chain-gene-cassettes
#14
Nam-Kyung Lee, Scott Bidlingmaier, Yang Su, Bin Liu
Monoclonal antibodies and antibody-derived therapeutics have emerged as a rapidly growing class of biological drugs for the treatment of cancer, autoimmunity, infection, and neurological diseases. To support the development of human antibodies, various display techniques based on antibody gene repertoires have been constructed over the last two decades. In particular, scFv-antibody phage display has been extensively utilized to select lead antibodies against a variety of target antigens. To construct a scFv phage display that enables efficient antibody discovery, and optimization, it is desirable to develop a system that allows modular assembly of highly diverse variable heavy chain and light chain (Vκ and Vλ) repertoires...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29116497/construction-of-human-immune-and-naive-scfv-libraries
#15
Jonas Kügler, Florian Tomszak, André Frenzel, Michael Hust
Antibody phage display is the most commonly used in vitro selection technology for the generation of human recombinant antibodies and has yielded thousands of useful antibodies for research, diagnostics, and therapy. The prerequisite for successful generation of antibodies using phage display is the construction of high-quality antibody gene libraries. Here, we give the detailed methods for the construction of human immune and naive scFv gene libraries.
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29100408/a-novel-fc-engineered-human-icam-1-cd54-antibody-with-potent-anti-myeloma-activity-developed-by-cellular-panning-of-phage-display-libraries
#16
Katja Klausz, Michael Cieker, Christian Kellner, Hans-Heinrich Oberg, Dieter Kabelitz, Thomas Valerius, Renate Burger, Martin Gramatzki, Matthias Peipp
To identify antibodies suitable for multiple myeloma (MM) immunotherapy, a cellular screening approach was developed using plasma cell lines JK-6L and INA-6 and human synthetic single-chain fragment variable (scFv) phage libraries. Isolated phage antibodies were screened for myeloma cell surface reactivity. Due to its binding characteristics, phage PIII-15 was selected to generate the scFv-Fc fusion protein TP15-Fc with an Fc domain optimized for FcγRIIIa binding. Various MM cell lines and patient-derived CD138-positive malignant plasma cells, but not granulocytes, B or T lymphocytes from healthy donors were recognized by TP15-Fc...
September 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/29089574/high-throughput-discovery-of-influenza-virus-neutralizing-antibodies-from-phage-displayed-synthetic-antibody-libraries
#17
Ing-Chien Chen, Yi-Kai Chiu, Chung-Ming Yu, Cheng-Chung Lee, Chao-Ping Tung, Yueh-Liang Tsou, Yi-Jen Huang, Chia-Lung Lin, Hong-Sen Chen, Andrew H-J Wang, An-Suei Yang
Pandemic and epidemic outbreaks of influenza A virus (IAV) infection pose severe challenges to human society. Passive immunotherapy with recombinant neutralizing antibodies can potentially mitigate the threats of IAV infection. With a high throughput neutralizing antibody discovery platform, we produced artificial anti-hemagglutinin (HA) IAV-neutralizing IgGs from phage-displayed synthetic scFv libraries without necessitating prior memory of antibody-antigen interactions or relying on affinity maturation essential for in vivo immune systems to generate highly specific neutralizing antibodies...
October 31, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29076991/single-chain-antibody-fragment-against-venom-from-the-snake-daboia-russelii-formosensis
#18
Chi-Hsin Lee, Yu-Ching Lee, Yueh-Lun Lee, Sy-Jye Leu, Liang-Tzung Lin, Chi-Ching Chen, Jen-Ron Chiang, Pharaoh Fellow Mwale, Bor-Yu Tsai, Ching-Sheng Hung, Yi-Yuan Yang
Russell's vipers containing hemotoxic and neurotoxic venom commonly cause snake envenomation. Horse-derived antivenom is a specific antidote, but its production is expensive and has side effects. Developing a cost-effective and more tolerable therapeutic strategy is favorable. In this study, using glutaraldehyde-attenuated Daboia russelii formosensis (DRF) venom proteins to immunize chickens, polyclonal yolk-immunoglobulin (IgY) antibodies were generated and showed a specific binding affinity. Phage display technology was used to generate two antibody libraries of single-chain variable fragments (scFvs) containing 3...
October 27, 2017: Toxins
https://www.readbyqxmd.com/read/29029420/generation-and-evaluation-of-antibody-agents-for-molecular-imaging-of-cd44v6-expressing-cancers
#19
Anna-Karin Haylock, Johan Nilvebrant, Anja Mortensen, Irina Velikyan, Marika Nestor, Ronny Falk
AIM: The aim of this study was to generate and characterize scFv antibodies directed to human CD44v6, as well as to radiolabel and evaluate top candidates in vitro and in vivo for their potential use in CD44v6-targeted molecular imaging in cancer patients. MATERIALS AND METHODS: Phage display selections were used to isolate CD44v6-specific scFvs. A chain shuffling strategy was employed for affinity maturation based on a set of CD44v6-specific first-generation clones...
September 12, 2017: Oncotarget
https://www.readbyqxmd.com/read/29018324/design-and-characterization-of-a-human-monoclonal-antibody-that-modulates-mutant-connexin-26-hemichannels-implicated-in-deafness-and-skin-disorders
#20
Liang Xu, Andrea Carrer, Francesco Zonta, Zhihu Qu, Peixiang Ma, Sheng Li, Federico Ceriani, Damiano Buratto, Giulia Crispino, Veronica Zorzi, Gaia Ziraldo, Francesca Bruno, Chiara Nardin, Chiara Peres, Flavia Mazzarda, Anna M Salvatore, Marcello Raspa, Ferdinando Scavizzi, Youjun Chu, Sichun Xie, Xuemei Yang, Jun Liao, Xiao Liu, Wei Wang, Shanshan Wang, Guang Yang, Richard A Lerner, Fabio Mammano
Background: Mutations leading to changes in properties, regulation, or expression of connexin-made channels have been implicated in 28 distinct human hereditary diseases. Eight of these result from variants of connexin 26 (Cx26), a protein critically involved in cell-cell signaling in the inner ear and skin. Lack of non-toxic drugs with defined mechanisms of action poses a serious obstacle to therapeutic interventions for diseases caused by mutant connexins. In particular, molecules that specifically modulate connexin hemichannel function without affecting gap junction channels are considered of primary importance for the study of connexin hemichannel role in physiological as well as pathological conditions...
2017: Frontiers in Molecular Neuroscience
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