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https://www.readbyqxmd.com/read/29119245/development-and-characterisation-of-a-novel-glucagon-like-peptide-1-receptor-antibody
#1
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/29118005/redirecting-t-cells-to-hematological-malignancies-with-bispecific-antibodies
#2
Mireya Paulina Velasquez, Challice L Bonifant, Stephen Gottschalk
There is a need to improve outcomes for patients with recurrent and/or refractory hematological malignancies. Immunotherapy holds the promise to meet this need since it does not rely on the cytotoxic mechanism of conventional therapies. Among different forms of immunotherapy, redirecting T cells to hematological malignancies with bispecific antibodies (BsAbs) is an attractive strategy. BsAbs are an 'off-the-shelf' product that is easily scalable in contrast to adoptive T-cell therapies. Among these, the bispecific T-cell engager (BiTE) blinatumomab has emerged as the most successful BsAb to date...
November 8, 2017: Blood
https://www.readbyqxmd.com/read/29116506/construction-of-chicken-antibody-libraries
#3
Jeanni Fehrsen, Susan Wemmer, Wouter van Wyngaardt
Recombinant antibody libraries based on chicken immunoglobulin genes are potentially valuable sources of phage displayed scFvs for use in veterinary diagnostics and research. The libraries described in this chapter are based on chicken variable heavy and light chain immunoglobulin genes joined by a short flexible linker cloned in the phagemid vector pHEN1. The resulting phagemids produce either scFvs displayed on the surface of the fusion phage subsequent to co-infection with helper phage, or soluble scFvs following IPTG induction...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29097914/cd44v6-targeted-imaging-of-head-and-neck-squamous-cell-carcinoma-antibody-based-approaches
#4
REVIEW
Diana Spiegelberg, Johan Nilvebrant
Head and neck squamous cell carcinoma (HNSCC) is a common and severe cancer with low survival rate in advanced stages. Noninvasive imaging of prognostic and therapeutic biomarkers could provide valuable information for planning and monitoring of the different therapy options. Thus, there is a major interest in development of new tracers towards cancer-specific molecular targets to improve diagnostic imaging and treatment. CD44v6, an oncogenic variant of the cell surface molecule CD44, is a promising molecular target since it exhibits a unique expression pattern in HNSCC and is associated with drug- and radio-resistance...
2017: Contrast Media & Molecular Imaging
https://www.readbyqxmd.com/read/29094051/improved-scfv-anti-lox-1-binding-activity-by-fusion-with-lox-1-binding-peptides
#5
Wei Hu, Qiuhong Xie, Hongyu Xiang
The oxidized low-density lipoprotein receptor-1 (LOX-1) targeted single-chain variable fragment (scFvs) is a promising molecule for the targeted delivery of imaging and therapeutic molecules of atherosclerotic diseases; however, its applications are limited by the inherent low antigen affinity. In this study, the three-dimensional (3D) model of the anti-LOX-1 scFv was constructed and its docking with the LOX-1 protein was developed. To improve the LOX-1-binding activity, the anti-LOX-1 scFv was designed to fuse with one of three LOX-1-binding heptapeptides, LTPATAI, FQTPPQL, and LSIPPKA, at its N-terminus and C-terminus and in the linker region, which have different LOX-1-binding interfaces with the anti-LOX-1 scFv analyzed by an array of computational approaches...
2017: BioMed Research International
https://www.readbyqxmd.com/read/29090224/production-and-evaluation-of-specific-single-chain-antibodies-against-ctla-4-for-cancer-targeted-therapy
#6
Farideh Hosseinzadeh, Saeed Mohammadi, Foroogh Nejatollahi
BACKGROUND: Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) molecules are expressed on T-cells and inhibit their function by inhibiting activation of subsequent T-cell molecular pathways. Blocking of CTLA-4 inhibits the growth of malignant tumor cells. Anti-CTLA-4 monoclonal antibodies activate the immune system against cancer. Due to several advantages of single-chain antibodies (scFvs) compared to monoclonal antibodies in cancer immunotherapy, specific anti-CTLA-4 scFvs (single-chain variable fragment) were selected in this study...
October 2017: Reports of Biochemistry & Molecular Biology
https://www.readbyqxmd.com/read/29076991/single-chain-antibody-fragment-against-venom-from-the-snake-daboia-russelii-formosensis
#7
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
#8
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/29027080/different-expression-systems-resulted-in-varied-binding-properties-of-anti-paclitaxel-single-chain-variable-fragment-antibody-clone-1c2
#9
Gorawit Yusakul, Seiichi Sakamoto, Poomraphie Nuntawong, Hiroyuki Tanaka, Satoshi Morimoto
The binding properties of recombinant antibody fragments, such as affinity and specificity, determine their usefulness for therapeutic and analytical applications. Anti-paclitaxel single-chain variable fragment clone 1C2 (anti-PT scFv1C2) was expressed using Escherichia coli cell and Bombyx mori larvae expression systems. The binding characteristics of the scFvs were evaluated using indirect competitive ELISA. The linear range of binding between anti-PT scFv1C2 and paclitaxel was significantly different between the anti-PT scFv1C2s derived from E...
October 12, 2017: Journal of Natural Medicines
https://www.readbyqxmd.com/read/28994732/production-of-single-chain-fv-antibodies-specific-for-ga-pyridine-an-advanced-glycation-end-product-age-with-reduced-inter-domain-motion
#10
Natsuki Fukuda, Kentaro Noi, Lidong Weng, Yoshihiro Kobashigawa, Hiromi Miyazaki, Yukari Wakeyama, Michiyo Takaki, Yusuke Nakahara, Yuka Tatsuno, Makiyo Uchida-Kamekura, Yoshiaki Suwa, Takashi Sato, Naoki Ichikawa-Tomikawa, Motoyoshi Nomizu, Yukio Fujiwara, Fumina Ohsaka, Takashi Saitoh, Katsumi Maenaka, Hiroyuki Kumeta, Shoko Shinya, Chojiro Kojima, Teru Ogura, Hiroshi Morioka
Due to their lower production cost compared with monoclonal antibodies, single-chain variable fragments (scFvs) have potential for use in several applications, such as for diagnosis and treatment of a range of diseases, and as sensor elements. However, the usefulness of scFvs is limited by inhomogeneity through the formation of dimers, trimers, and larger oligomers. The scFv protein is assumed to be in equilibrium between the closed and open states formed by assembly or disassembly of VH and VL domains. Therefore, the production of an scFv with equilibrium biased to the closed state would be critical to overcome the problem in inhomogeneity of scFv for industrial or therapeutic applications...
October 10, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28981720/selection-of-novel-affinity-matured-human-chondroitin-sulfate-proteoglycan-4-antibody-fragments-by-yeast-display
#11
Xin Yu, Liang Qu, Darell D Bigner, Vidyalakshmi Chandramohan
Chondroitin sulfate proteoglycan 4 (CSPG4) is a promising target for cancer immunotherapy due to its high level of expression in a number of malignant tumors, and its essential role in tumor growth and progression. Clinical application of CSPG4-targeting immunotherapies is hampered by the lack of fully human high-affinity CSPG4 antibodies or antibody fragments. To overcome this limitation, we performed affinity maturation on a novel human CSPG4 single-chain Fv fragment (scFv) using the random mutagenesis approach and screened for improved variants from a yeast display library using a modified whole-cell panning method followed by fluorescence-activated cell sorting...
September 1, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28942640/development-of-an-indirect-competitive-enzyme-linked-immunosorbent-assay-for-glycocholic-acid-based-on-chicken-single-chain-variable-fragment-antibodies
#12
Xiping Cui, Natalia Vasylieva, Panpan Wu, Bogdan Barnych, Jun Yang, Ding Shen, Qiyi He, Shirley J Gee, Suqing Zhao, Bruce D Hammock
Glycocholic acid (GCA) is an important metabolite of bile acids, whose urine levels are expected to be a specific diagnostic biomarker for hepatocellular carcinoma (HCC). A high-throughput immunoassay for determination of GCA would be of significant advantage and useful for primary diagnosis, surveillance, and early detection of HCC. Single-chain variable fragment (scFv) antibodies have several desirable characteristics and are an attractive alternative to traditional antibodies for the immunoassay. Because chicken antibodies possess single heavy and light variable functional domains, they are an ideal framework for simplified generation of recombinant antibodies for GCA detection...
October 17, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28938888/unique-binding-modes-for-the-broad-neutralizing-activity-of-single-chain-variable-fragments-scfv-targeting-cd4-induced-epitopes
#13
Kazuki Tanaka, Takeo Kuwata, Muntasir Alam, Gilad Kaplan, Shokichi Takahama, Kristel Paola Ramirez Valdez, Anna Roitburd-Berman, Jonathan M Gershoni, Shuzo Matsushita
BACKGROUND: The CD4-induced (CD4i) epitopes in gp120 includes the co-receptor binding site, which are formed and exposed after interaction with CD4. Monoclonal antibodies (mAbs) to the CD4i epitopes exhibit limited neutralizing activity because of restricted access to their epitopes. However, small fragment counterparts such as single-chain variable fragments (scFvs) have been reported to neutralize a broad range of viruses compared with the full-size IgG molecule. To identify the CD4i epitope site responsible for this broad neutralization we constructed three scFvs of anti-CD4i mAbs from a human immunodeficiency virus type 1 (HIV-1)-infected elite controller, and investigated the neutralization coverage and precise binding site in the CD4i epitopes...
September 22, 2017: Retrovirology
https://www.readbyqxmd.com/read/28919896/vhh-based-bispecific-antibodies-targeting-cytokine-production
#14
Maxim A Nosenko, Kamar-Sulu N Atretkhany, Vladislav V Mokhonov, Grigory A Efimov, Andrey A Kruglov, Sergei V Tillib, Marina S Drutskaya, Sergei A Nedospasov
Proinflammatory cytokines, such as TNF, IL-6, and IL-1, play pathogenic roles in multiple diseases and are attractive targets for biologic drugs. Because proinflammatory cytokines possess non-redundant protective and immunoregulatory functions, their systemic neutralization carries the potential for unwanted side effects. Therefore, next-generation anti-cytokine therapies would seek to selectively neutralize pathogenic cytokine signaling, leaving normal function intact. Fortunately, the biology of proinflammatory cytokines provides several such opportunities...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28900185/anti-a%C3%AE-single-chain-variable-fragment-antibodies-restore-memory-acquisition-in-a-drosophila-model-of-alzheimer-s-disease
#15
Alfonso Martin-Peña, Diego E Rincon-Limas, Pedro Fernandez-Funez
Alzheimer's disease (AD) is a prevalent neurodegenerative disorder triggered by the accumulation of soluble assemblies of the amyloid-β42 (Aβ42) peptide. Despite remarkable advances in understanding the pathogenesis of AD, the development of palliative therapies is still lacking. Engineered anti-Aβ42 antibodies are a promising strategy to stall the progression of the disease. Single-chain variable fragment (scFv) antibodies increase brain penetration and offer flexible options for delivery while maintaining the epitope targeting of full antibodies...
September 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28888872/the-catalytic-activity-of-a-recombinant-single-chain-variable-fragment-nucleic-acid-hydrolysing-antibody-varies-with-fusion-tag-and-expression-host
#16
Joungmin Lee, Minjae Kim, Youngsil Seo, Yeonjin Lee, Hyunjoon Park, Sung June Byun, Myung-Hee Kwon
The antigen-binding properties of single chain Fv antibodies (scFvs) can vary depending on the position and type of fusion tag used, as well as the host cells used for expression. The issue is even more complicated with a catalytic scFv antibody that binds and hydrolyses a specific antigen. Herein, we investigated the antigen-binding and -hydrolysing activities of the catalytic anti-nucleic acid antibody 3D8 scFv expressed in Escherichia coli or HEK293f cells with or without additional amino acid residues at the N- and C-termini...
November 1, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28887121/discovery-of-human-scfvs-that-cross-neutralize-the-toxic-effects-of-b-jararacussu-and-c-d-terrificus-venoms
#17
Luciano C Silva, Manuela B Pucca, Gabriela Pessenda, Lucas B Campos, Edson Z Martinez, Felipe A Cerni, José E Barbosa
Accidents involving venomous snakes are a public health problem worldwide, causing a large number of deaths per year. In Brazil, the majority of accidents are caused by the Bothrops and Crotalus genera, which are responsible for approximately 80% of severe envenoming cases. The cross-neutralization of snake venoms by antibodies is an important issue for development of more effective treatments. Our group has previously reported the construction of human monoclonal antibody fragments towards Bothrops jararacussu and Crotalus durissus terrificus' venoms...
September 5, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28860479/de-novo-design-of-antibody-complementarity-determining-regions-binding-a-flag-tetra-peptide
#18
Kevin C Entzminger, Jeong-Min Hyun, Robert J Pantazes, Athena C Patterson-Orazem, Ahlam N Qerqez, Zach P Frye, Randall A Hughes, Andrew D Ellington, Raquel L Lieberman, Costas D Maranas, Jennifer A Maynard
Computational antibody engineering efforts to date have focused on improving binding affinities or biophysical characteristics. De novo design of antibodies binding specific epitopes could greatly accelerate discovery of therapeutics as compared to conventional immunization or synthetic library selection strategies. Here, we employed de novo complementarity determining region (CDR) design to engineer targeted antibody-antigen interactions using previously described in silico methods. CDRs predicted to bind the minimal FLAG peptide (Asp-Tyr-Lys-Asp) were grafted onto a single-chain variable fragment (scFv) acceptor framework...
August 31, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28846502/rare-high-affinity-anti-pathogen-antibodies-from-human-repertoires-discovered-using-microfluidics-and-molecular-genomics
#19
Adam S Adler, Rena A Mizrahi, Matthew J Spindler, Matthew S Adams, Michael A Asensio, Robert C Edgar, Jackson Leong, Renee Leong, Lucy Roalfe, Rebecca White, David Goldblatt, David S Johnson
Affinity-matured, functional anti-pathogen antibodies are present at low frequencies in natural human repertoires. These antibodies are often excellent candidates for therapeutic monoclonal antibodies. However, mining natural human antibody repertoires is a challenge. In this study, we demonstrate a new method that uses microfluidics, yeast display, and deep sequencing to identify 247 natively paired anti-pathogen single-chain variable fragments (scFvs), which were initially as rare as 1 in 100,000 in the human repertoires...
November 2017: MAbs
https://www.readbyqxmd.com/read/28846130/identification-and-characterisation-of-the-proteins-bound-by-specific-phage-displayed-recombinant-antibodies-scfv-obtained-against-brazil-nut-and-almond-extracts
#20
Silvia de la Cruz, Raquel Madrid, Aina García-García, Marcos Alcocer, Rosario Martín, Isabel González, Teresa García
BACKGROUND: Almonds and Brazil nuts are widely consumed allergenic nuts whose presence must be declared according to food labelling regulations. Their detection in food products has been recently achieved by ELISA methods with recombinant antibodies (scFv) isolated against complete Brazil nut and almond protein extracts. The screening of phage-scFv libraries against complete protein extracts confers a series of advantages over the use of purified proteins, as recombinant proteins might alter their native folding...
August 28, 2017: Journal of the Science of Food and Agriculture
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