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Antibody engineering

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https://www.readbyqxmd.com/read/28342851/site-specific-photocoupling-of-pbpa-mutated-scfv-antibodies-for-use-in-affinity-proteomics
#1
Mattias Brofelth, Lars Wagner Städe, Anna Isinger Ekstrand, Linn Petersson Edfeldt, Rebeka Kovačič, Thorbjørn Terndrup Nielsen, Kim Lambertsen Larsen, Laurent Duroux, Christer Wingren
Recombinant antibody libraries can provide a source of renewable and high-performing binders tailored for use in affinity proteomics. In this context, the process of generating site-specific 1:1 tagging/functionalization and/or orientated surface immobilization of antibodies has, however, proved to be challenging. Hence, novel ways of generating such engineered antibodies for use in affinity proteomics could have a major impact on array performance. In this study, we have further tailored the design of human recombinant scFv antibodies for site-specific photocoupling through the use of an unnatural amino acid (UAA) and the Dock'n'Flash technology...
March 22, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28336959/affibody-molecules-as-engineered-protein-drugs
#2
REVIEW
Fredrik Y Frejd, Kyu-Tae Kim
Affibody molecules can be used as tools for molecular recognition in diagnostic and therapeutic applications. There are several preclinical studies reported on diagnostic and therapeutic use of this molecular class of alternative scaffolds, and early clinical evidence is now beginning to accumulate that suggests the Affibody molecules to be efficacious and safe in man. The small size and ease of engineering make Affibody molecules suitable for use in multispecific constructs where AffiMabs is one such that offers the option to potentiate antibodies for use in complex disease...
March 24, 2017: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/28330604/stable-and-potent-selenomab-drug-conjugates
#3
Xiuling Li, Christopher G Nelson, Rajesh R Nair, Lori Hazlehurst, Tina Moroni, Pablo Martinez-Acedo, Alex R Nanna, David Hymel, Terrence R Burke, Christoph Rader
Selenomabs are engineered monoclonal antibodies with one or more translationally incorporated selenocysteine residues. The unique reactivity of the selenol group of selenocysteine permits site-specific conjugation of drugs. Compared with other natural and unnatural amino acid and carbohydrate residues that have been used for the generation of site-specific antibody-drug conjugates, selenocysteine is particularly reactive, permitting fast, single-step, and efficient reactions under near physiological conditions...
March 9, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/28330372/car-t-cell-therapy-progress-and-prospects
#4
Olivia Wilkins, Allison May Keeler, Terence R Flotte
Lentivirus-mediated transduction of autologous T-cells with a chimeric antigen receptor (CAR) to confer a desired epitope-specificity as a targeted immunotherapy for cancer has been among the first human gene therapy techniques to demonstrate widespread therapeutic efficacy. Other approaches to using gene therapy to enhance anti-tumor immunity have been less specific and less effective. These included amplification, marking, and cytokine transduction of tumor infiltrating lymphocytes (TIL), recombinant virus-based expression of tumor antigens as a tumor vaccine, and transduction of antigen-presenting cells (APCs) with tumor antigens...
March 23, 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28325297/engineering-of-pedf-expressing-primary-pigment-epithelial-cells-by-the-sb-transposon-system-delivered-by-pfar4-plasmids
#5
Gabriele Thumann, Nina Harmening, Cécile Prat-Souteyrand, Corinne Marie, Marie Pastor, Attila Sebe, Csaba Miskey, Laurence D Hurst, Sabine Diarra, Martina Kropp, Peter Walter, Daniel Scherman, Zoltán Ivics, Zsuzsanna Izsvák, Sandra Johnen
Neovascular age-related macular degeneration (nvAMD) is characterized by choroidal blood vessels growing into the subretinal space, leading to retinal pigment epithelial (RPE) cell degeneration and vision loss. Vessel growth results from an imbalance of pro-angiogenic (e.g., vascular endothelial growth factor [VEGF]) and anti-angiogenic factors (e.g., pigment epithelium-derived factor [PEDF]). Current treatment using intravitreal injections of anti-VEGF antibodies improves vision in about 30% of patients but may be accompanied by side effects and non-compliance...
March 17, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28325214/antibody-based-cancer-therapy-successful-agents-and-novel-approaches
#6
D Hendriks, G Choi, M de Bruyn, V R Wiersma, E Bremer
Since their discovery, antibodies have been viewed as ideal candidates or "magic bullets" for use in targeted therapy in the fields of cancer, autoimmunity, and chronic inflammatory disorders. A wave of antibody-dedicated research followed, which resulted in the clinical approval of a first generation of monoclonal antibodies for cancer therapy such as rituximab (1997) and cetuximab (2004), and infliximab (2002) for the treatment of autoimmune diseases. More recently, the development of antibodies that prevent checkpoint-mediated inhibition of T cell responses invigorated the field of cancer immunotherapy...
2017: International Review of Cell and Molecular Biology
https://www.readbyqxmd.com/read/28322923/microspheres-prime-rmva-boost-vaccination-enhances-humoral-and-cellular-immune-response-in-ifnar-mice-conferring-protection-against-serotypes-1-and-4-of-bluetongue-virus
#7
Alejandro Marín-López, Eva Calvo-Pinilla, Diego Barriales, Gema Lorenzo, Javier Benavente, Alejandro Brun, Jose Manuel Martínez-Costas, Javier Ortego
Bluetongue virus (BTV) is the causative agent of bluetongue disease (BT), which affects domestic and wild ruminants. At the present, 27 different serotypes have been documented. Vaccination has been demonstrated as one of the most effective methods to avoid viral dissemination. To overcome the drawbacks associated with the use of inactivated and attenuated vaccines we engineered a new recombinant BTV vaccine candidate based on proteins VP2, VP7, and NS1 of BTV-4 that were incorporated into avian reovirus muNS-Mi microspheres (MS-VP2/VP7/NS1) and recombinant modified vaccinia virus Ankara (rMVA)...
March 16, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28322517/materials-for-microfluidic-immunoassays-a-review
#8
REVIEW
Lei Mou, Xingyu Jiang
Conventional immunoassays suffer from at least one of these following limitations: long processing time, high costs, poor user-friendliness, technical complexity, poor sensitivity and specificity. Microfluidics, a technology characterized by the engineered manipulation of fluids in channels with characteristic lengthscale of tens of micrometers, has shown considerable promise for improving immunoassays that could overcome these limitations in medical diagnostics and biology research. The combination of microfluidics and immunoassay can detect biomarkers with faster assay time, reduced volumes of reagents, lower power requirements, and higher levels of integration and automation compared to traditional approaches...
March 21, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28320976/a-study-on-the-efficacy-of-the-recombinant-yersinia-adhesin-a-vaccine-against-yersiniosis-in-the-early-phase
#9
Kosuke Tsugo, Shin/Ichi Nakamura, Hiroko Yamanaka, Yumi Une
Yersinia pseudotuberculosis (Y. ptb) is a zoonotic pathogenic bacterial species of the family Enterobacteriaceae and causes yersiniosis, an acute intestinal infection in humans and animals. Y. ptb is often implicated in lethal epidemics in zoo animals and reductions in the breeding population, but a valid prevention method has not been established. Therefore, this study aimed to develop a vaccine for yersiniosis control. The immunogenicity of one of the adhesion factors involved in pathogenic mechanisms of Y...
March 20, 2017: Journal of Veterinary Medical Science
https://www.readbyqxmd.com/read/28319663/2h-ws2-quantum-dots-produced-by-modulating-the-dimension-and-phase-of-1t-nanosheets-for-antibody-free-optical-sensing-of-neurotransmitters
#10
Man-Jin Kim, Su-Ji Jeon, Tae Woog Kang, Jong-Min Ju, DaBin Yim, Hye-In Kim, Jung Hyun Park, Jong-Ho Kim
Modulating the dimensions and phases of transition metal dichalcogenides is of great interest to enhance their intrinsic properties or to create new physicochemical properties. Herein, we report an effective approach to synthesize 2H-WS2 quantum dots (QDs) via the dimension and phase engineering of 1T-WS2 nanosheets. The solvothermal reaction of chemically-exfoliated 1T-WS2 nanosheets in N-methyl-2-pyrrolidone (NMP) under an N2 atmosphere induced their chopping and phase transition at lower temperature to produce 2H-WS2 QDs with a high quantum yield (5...
March 20, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28319021/engineering-a-growth-sensor-to-select-intracellular-antibodies-in-the-cytosol-of-mammalian-cells
#11
Thuy Duong Nguyen, Hitoshi Takasuka, Yoshihiro Kaku, Satoshi Inoue, Teruyuki Nagamune, Masahiro Kawahara
Intracellular antibodies (intrabodies) are expected to function as therapeutics as well as tools for elucidating in vivo function of proteins. In this study, we propose a novel intrabody selection method in the cytosol of mammalian cells by utilizing a growth signal, induced by the interaction of the target antigen and an scFv-c-kit growth sensor. Here, we challenge this method to select specific intrabodies against rabies virus nucleoprotein (RV-N) for the first time. As a result, we successfully select antigen-specific intrabodies from a naïve synthetic library using phage panning followed by our growth sensor-based intracellular selection method, demonstrating the feasibility of the method...
March 16, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28318221/crystal-structure-of-a-homogeneous-igg-fc-glycoform-with-the-n-glycan-designed-to-maximize-the-antibody-dependent-cellular-cytotoxicity
#12
Chia-Lin Chen, Jen-Chi Hsu, Chin-Wei Lin, Chia-Hung Wang, Ming-Hung Tsai, Chung-Yi Wu, Chi-Huey Wong, Che Ma
N-glycosylation on IgG modulates Fc conformation and effector functions. An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen, but also in some cases provides gain-of-function effector activity. We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain, and solved its crystal structure...
March 20, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28316596/the-efficacy-of-humanized-antibody-against-the-sporothrix-antigen-gp70-in-promoting-phagocytosis-and-reducing-disease-burden
#13
José R F de Almeida, Karla L Santiago, Gilberto H Kaihami, Andrea Q Maranhão, Marcelo de Macedo Brígido, Sandro R de Almeida
Sporotrichosis is a subcutaneous mycosis distributed worldwide and is frequently reported in countries with tropical climates, as Latin America countries. We previously demonstrated that mice with sporotrichosis produce specific antibodies against a 70-kDa fungal protein, indicating that specific antibodies against this molecule may help to control the sporotrichosis. IgG1 monoclonal antibody was generated, and called mAbP6E7, in mice against a 70-kDa glycoprotein (gp70) of S. schenckii. The mAbP6E7 showed prophylactic and therapeutic activity against sporotrichosis...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28315359/antibodies-and-associates-partners-in-targeted-drug-delivery
#14
REVIEW
Patrick J Kennedy, Carla Oliveira, Pedro L Granja, Bruno Sarmento
Monoclonal antibodies (mAbs) are well established in the clinic due to their specificity and affinity to a diverse array of biochemical targets. More recently, mAbs are being exploited as targeting agents in modern drug delivery systems, aiming to bypass normal host tissue and to accumulate a therapeutic agent to a specific tissue or cell for enhanced pharmacology. At sizes ranging from ~10-100nm, antibody-based bioconjugates have opened up a whole new realm of clinical possibilities with several platforms emerging on the market...
March 14, 2017: Pharmacology & Therapeutics
https://www.readbyqxmd.com/read/28315215/detection-of-food-allergens-by-phage-displayed-produced-antibodies
#15
Raquel Madrid, Silvia de la Cruz, Aina García, Rosario Martín, Isabel González, Teresa García
Phage display is a powerful tool to produce recombinant antibodies against a given antigen without animal immunization. This technology employs libraries of recombinant bacteriophages that display billions of different functional antibody fragments on their surface. They are selected by panning in vitro against the target antigen in search for specific binders. In this chapter, we describe the selection of single chain variable fragment (scFv) antibodies to be used for detection of allergenic proteins from nuts in food products...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28303661/immunological-and-physiological-observations-in-baboons-with-life-supporting-genetically-engineered-pig-kidney-grafts
#16
Hayato Iwase, Hidetaka Hara, Mohamed Ezzelarab, Tao Li, Zhongqiang Zhang, Bingsi Gao, Hong Liu, Cassandra Long, Yi Wang, Amy Cassano, Edwin Klein, Carol Phelps, David Ayares, Abhinav Humar, Martin Wijkstrom, David K C Cooper
BACKGROUND: Genetically engineered pigs could provide a source of kidneys for clinical transplantation. The two longest kidney graft survivals reported to date have been 136 and 310 days, but graft survival >30 days has been unusual until recently. METHODS: Donor pigs (n=4) were on an α1,3-galactosyltransferase gene-knockout (GTKO)/human complement regulatory protein (CD46) background (GTKO/CD46). In addition, the pigs were transgenic for at least one human coagulation regulatory protein...
March 17, 2017: Xenotransplantation
https://www.readbyqxmd.com/read/28303026/strategies-and-challenges-for-the-next-generation-of-antibody-drug-conjugates
#17
REVIEW
Alain Beck, Liliane Goetsch, Charles Dumontet, Nathalie Corvaïa
Antibody-drug conjugates (ADCs) are one of the fastest growing classes of oncology therapeutics. After half a century of research, the approvals of brentuximab vedotin (in 2011) and trastuzumab emtansine (in 2013) have paved the way for ongoing clinical trials that are evaluating more than 60 further ADC candidates. The limited success of first-generation ADCs (developed in the early 2000s) informed strategies to bring second-generation ADCs to the market, which have higher levels of cytotoxic drug conjugation, lower levels of naked antibodies and more-stable linkers between the drug and the antibody...
March 17, 2017: Nature Reviews. Drug Discovery
https://www.readbyqxmd.com/read/28300660/mechanistic-understanding-of-the-cysteine-capping-modifications-of-antibodies-enables-selective-chemical-engineering-in-live-mammalian-cells
#18
Xiaotian Zhong, Tao He, Amar S Prashad, Wenge Wang, Justin Cohen, Darren Ferguson, Amy S Tam, Eric Sousa, Laura Lin, Lioudmila Tchistiakova, Scott Gatto, Aaron D'Antona, Yen-Tung Luan, Weijun Ma, Richard Zollner, Jing Zhou, Bo Arve, Will Somers, Ronald Kriz
Protein modifications by intricate cellular machineries often redesign the structure and function of existing proteins to impact biological networks. Disulfide bond formation between cysteine (Cys) pairs is one of the most common modifications found in extracellularly-destined proteins, key to maintaining protein structure. Unpaired surface cysteines on secreted mammalian proteins are also frequently found disulfide-bonded with free Cys or glutathione (GSH) in circulation or culture, the mechanism for which remains unknown...
March 11, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28299901/dual-display-of-proteins-on-the-yeast-cell-surface-simplifies-quantification-of-binding-interactions-and-enzymatic-bioconjugation-reactions
#19
Sungwon Lim, Jeff E Glasgow, Maria Filsinger Interrante, Erica M Storm, Jennifer R Cochran
Yeast surface display, a well-established technology for protein analysis and engineering, involves expressing a protein of interest as a genetic fusion to either the N- or C-terminus of the yeast Aga2p mating protein. Historically, yeast-displayed protein variants are flanked by peptide epitope tags that enable flow cytometric measurement of construct expression using fluorescent primary or secondary antibodies. Here, we built upon this technology to develop a new yeast display strategy that comprises fusion of two different proteins to Aga2p, one to the N-terminus and one to the C-terminus...
March 16, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28296934/reverse-lectin-elisa-for-detecting-fucosylated-forms-of-%C3%AE-1-acid-glycoprotein-associated-with-hepatocellular-carcinoma
#20
Eva Åström, Per Stål, Robin Zenlander, Pia Edenvik, Catharina Alexandersson, Mats Haglund, Ingvar Rydén, Peter Påhlsson
Altered fucosylation of glycoproteins is associated with development of hepatocellular carcinoma (HCC). Lectins have been commonly used to assay changes in fucosylation of plasma glycoproteins. In the present study a recombinantly engineered form of the fucose binding lectin Aleuria aurantia (AAL) consisting of a single binding site for fucose (S2), was used to construct a reverse lectin ELISA method. Microtiter plates coated with the S2 lectin were used to capture glycoproteins from plasma samples followed by antibody detection of S2-bound fucosylated α1-acid glycoprotein (S2-bound AGP)...
2017: PloS One
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