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https://www.readbyqxmd.com/read/28230642/strategies-to-obtain-diverse-and-specific-human-monoclonal-antibodies-from-transgenic-animals
#1
Marianne Brüggemann, Michael J Osborn, Biao Ma, Roland Buelow
Techniques to obtain large quantities of antigen-specific monoclonal antibodies, mAbs, were first established in the 1970s when Georges Köhler and César Milstein immortalized antibody-producing mouse B-lymphocytes by fusion with myeloma cells (http://www.whatisbiotechnology.org/exhibitions/milstein). Combined with the expression of human antibodies in transgenic animals, this technique allowed upon immunization the generation of highly specific fully human mAbs for therapeutic applications. Apart from being extremely beneficial, mAbs are a huge success commercially...
February 23, 2017: Transplantation
https://www.readbyqxmd.com/read/28228285/mass-spectrometry-profiling-of-hla-associated-peptidomes-in-mono-allelic-cells-enables-more-accurate-epitope-prediction
#2
Jennifer G Abelin, Derin B Keskin, Siranush Sarkizova, Christina R Hartigan, Wandi Zhang, John Sidney, Jonathan Stevens, William Lane, Guang Lan Zhang, Thomas M Eisenhaure, Karl R Clauser, Nir Hacohen, Michael S Rooney, Steven A Carr, Catherine J Wu
Identification of human leukocyte antigen (HLA)-bound peptides by liquid chromatography-tandem mass spectrometry (LC-MS/MS) is poised to provide a deep understanding of rules underlying antigen presentation. However, a key obstacle is the ambiguity that arises from the co-expression of multiple HLA alleles. Here, we have implemented a scalable mono-allelic strategy for profiling the HLA peptidome. By using cell lines expressing a single HLA allele, optimizing immunopurifications, and developing an application-specific spectral search algorithm, we identified thousands of peptides bound to 16 different HLA class I alleles...
February 21, 2017: Immunity
https://www.readbyqxmd.com/read/28223164/exploring-sensitivity-throughput-of-a-parallel-flow-spri-biosensor-for-characterization-of-antibody-antigen-interaction
#3
Vishal Kamat, Ashique Rafique
Rapid growth in the field of biotherapeutics has led to an increased demand for high-throughput, label-free biosensors exhibiting high sensitivity. To support the current needs, Sierra Sensors introduced a surface plasmon resonance imaging (SPRi) based biosensor, Molecular Affinity Screening System (MASS-1). We assessed the potential utility of MASS-1 to support Regeneron's therapeutic antibody discovery. A large panel of antibody-antigen interactions was characterized using MASS-1 and the kinetic data were compared with the Biacore 4000 biosensor...
February 18, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28218509/critical-role-of-inter-domain-interactions-on-the-conformational-change-and-catalytic-mechanism-of-endoplasmic-reticulum-aminopeptidase-1
#4
Athanasios Stamogiannos, Zachary Maben, Athanasios Papakyriakou, Anastasia Mpakali, Paraskevi Kokkala, Dimitris Georgiadis, Lawrence J Stern, Efstratios Stratikos
Endoplasmic Reticulum Aminopeptidase 1 (ERAP1) is an intracellular enzyme important for the generation of antigenic epitopes and class I MHC-restricted adaptive immune responses. ERAP1 processes a vast variety of different peptides but still shows length and sequence selectivity although the mechanism behind these properties is poorly understood. X-ray crystallographic analysis has revealed that ERAP1 can assume at least two distinct conformations in which the C-terminal domain IV is either proximal or distal to the active site domain II...
February 20, 2017: Biochemistry
https://www.readbyqxmd.com/read/28213228/smartphone-based-immunosensor-for-ca125-detection
#5
Oana Hosu, Andrea Ravalli, Giuseppe Mattia Lo Piccolo, Cecilia Cristea, Robert Sandulescu, Giovanna Marrazza
In this work, we report the design, the development and the characterization of the analytical performances of a colorimetric smartphone-based immunosensor for the detection of cancer antigen 125 (CA125). The immunosensor was based on a sandwich strategy in which the primary antibody was immobilized by spotting onto the 3D nitrocellulose membrane. The immunospots were subsequently incubated with CA125 solutions, followed by the affinity reaction with a secondary antibody labeled with gold nanoparticles (AuNPs)...
May 1, 2017: Talanta
https://www.readbyqxmd.com/read/28211613/novel-anticarcinoembryonic-antigen-antibody-drug-conjugate-has-antitumor-activity-in-the-existence-of-soluble-antigen
#6
Daisuke Shinmi, Ryosuke Nakano, Keisuke Mitamura, Minami Suzuki-Imaizumi, Junko Iwano, Yuya Isoda, Junichi Enokizono, Yasuhisa Shiraishi, Emi Arakawa, Kazuma Tomizuka, Kazuhiro Masuda
Carcinoembryonic antigen (CEA) is a classic tumor-specific antigen that is overexpressed in several cancers, including gastric cancer. Although some anti-CEA antibodies have been tested, to the best of our knowledge, there are currently no clinically approved anti-CEA antibody therapies. Because of this, we have created the novel anti-CEA antibody, 15-1-32, which exhibits stronger binding to membrane-bound CEA on cancer cells than existing anti-CEA antibodies. 15-1-32 also shows poor affinity for soluble CEA; thus, the binding activity of 15-1-32 to membrane-bound CEA is not influenced by soluble CEA...
February 17, 2017: Cancer Medicine
https://www.readbyqxmd.com/read/28211542/identification-of-the-core-regulators-of-the-hla-i-peptide-binding-process
#7
Yu-Hang Zhang, Zhihao Xing, Chenglin Liu, ShaoPeng Wang, Tao Huang, Yu-Dong Cai, Xiangyin Kong
During the display of peptide/human leukocyte antigen (HLA) -I complex for further immune recognition, the cleaved and transported antigenic peptides have to bind to HLA-I protein and the binding affinity between peptide epitopes and HLA proteins directly influences the immune recognition ability in human beings. Key factors affecting the binding affinity during the generation, selection and presentation processes of HLA-I complex have not yet been fully discovered. In this study, a new method describing the HLA class I-peptide interactions was proposed...
February 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28207486/particle-based-delivery-of-the-hiv-envelope-protein
#8
Benedikt Asbach, Ralf Wagner
PURPOSE OF REVIEW: A major focus in HIV vaccine research is the development of suitable antigens that elicit broadly neutralizing antibody responses targeting HIV's envelope protein (Env). Delivery of Env in a repetitive manner on particle-based carriers allows higher avidity interactions and is therefore expected to efficiently engage B cells, thus leading to affinity maturation that results in superior antibody responses characterized by improved breadth, potency, and durability. This review summarizes current work that is evaluating diverse types of such particulate carriers for Env delivery...
February 15, 2017: Current Opinion in HIV and AIDS
https://www.readbyqxmd.com/read/28202413/engineered-antibody-ch2-domains-binding-to-nucleolin-isolation-characterization-and-improvement-of-aggregation
#9
De-Zhi Li, Rui Gong, Jun Zheng, Xi-Hai Chen, Dimiter S Dimitrov, Qi Zhao
Smaller recombinant antibody fragments are now emerging as alternatives of conventional antibodies. Especially, immunoglobulin (Ig) constant CH2 domain and engineered CH2 with improved stability are promising as scaffolds for selection of specific binders to various antigens. We constructed a yeast display library based on an engineered human IgG1 CH2 scaffold with diversified loop regions. A group of CH2 binders were isolated from this yeast display library by panning against nucleolin, which is a tumor-associated antigen involved in cell proliferation, tumor cell growth and angiogenesis...
February 12, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28195200/dynamical-footprint-of-cross-reactivity-in-a-human-autoimmune-t-cell-receptor
#10
Amit Kumar, Francesco Delogu
The present work focuses on the dynamical aspects of cross-reactivity between myelin based protein (MBP) self-peptide and two microbial peptides (UL15, PMM) for Hy.1B11 T-cell receptor (TCR). This same TCR was isolated from a patient suffering from multiple sclerosis (MS). The study aims at highlighting the chemical interactions underlying recognition mechanisms between TCR and the peptides presented by Major Histocompatibility Complex (MHC) proteins, which form a crucial component in adaptive immune response against foreign antigens...
February 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28193211/display-of-single-chain-variable-fragments-on-bacteriophage-ms2-virus-like-particles
#11
Christopher A Lino, Jerri C Caldeira, David S Peabody
BACKGROUND: Virus-like particles (VLPs) of the RNA bacteriophage MS2 have many potential applications in biotechnology. MS2 VLPs provide a platform for peptide display and affinity selection (i.e. biopanning). They are also under investigation as vehicles for targeted drug delivery, using display of receptor-specific peptides or nucleic acid aptamers to direct their binding to specific cell-surface receptors. However, there are few molecules more suited to the precise targeting and binding of a cellular receptor than antibodies...
February 13, 2017: Journal of Nanobiotechnology
https://www.readbyqxmd.com/read/28193005/mapping-the-energetic-epitope-of-an-antibody-interleukin-23-interaction-with-hydrogen-deuterium-exchange-fast-photochemical-oxidation-of-proteins-mass-spectrometry-and-alanine-shave-mutagenesis
#12
Jing Li, Hui Wei, Stanley R Krystek, Derek Bond, Ty M Brender, Daniel Cohen, Jena Feiner, Nels Hamacher, Johanna Harshman, Richard Y-C Huang, Susan H Julien, Zheng Lin, Kristina Moore, Luciano Mueller, Claire Noriega, Preeti Sejwal, Paul Sheppard, Brenda Stevens, Guodong Chen, Adrienne A Tymiak, Michael L Gross, Lumelle A Schneeweis
Epitope mapping the specific residues of an antibody/antigen interaction can be used to support mechanistic interpretation, antibody optimization, and epitope novelty assessment. Thus, there is a strong need for mapping methods, particularly integrative ones. Here, we report the identification of an energetic epitope by determining the interfacial hot-spot that dominates the binding affinity for an anti-interleukin-23 (anti-IL-23) antibody by using the complementary approaches of hydrogen/deuterium exchange mass spectrometry (HDX-MS), fast photochemical oxidation of proteins (FPOP), alanine shave mutagenesis, and binding analytics...
February 9, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28192940/analysis-of-tertiary-interactions-between-sart3-and-u6-small-nuclear-rna-using-modified-nanocapillaries
#13
Choongman Lee, Joon Kyu Park, Yeoan Youn, Joo Hyoung Kim, Kyo-Seok Lee, Nak-Kyoon Kim, Eunji Kim, Eunice Eunkyeong Kim, Kyung-Hwa Yoo
We employed modified glass nanocapillaries to investigate interactions between the RNA-binding protein, known as cell carcinoma antigen recognized by T cells-3 (SART3), and the noncoding spliceosome component, U6 small nuclear RNA (snRNA), at the single-molecule level. We functionalized the nanocapillaries with U6 snRNA fragments, which were hybridized to DNA molecules and then covalently attached to the nanocapillary surface. When transported through the modified nanocapillaries, two different SART3-derived constructs, HAT-RRM1-RRM2 and RRM1-RRM2, exhibited resistive ionic current pulses with different dwell times, which represented their different binding affinities to tethered U6 snRNAs...
February 9, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28190897/synthesis-of-multivalent-glycopeptide-conjugates-that-mimic-an-hiv-epitope
#14
Jennifer K Bailey, Dung N Nguyen, Satoru Horiya, Isaac J Krauss
Recently, we reported a directed evolution method which enabled us to discover sequences of glycopeptides that bind with picomolar affinity to HIV antibody 2G12 and are of interest as HIV vaccine candidates. In this manuscript, we describe the syntheses of several of these large (~11-12 kDa) glycopeptides by a combination of fast flow peptide synthesis and click chemistry. We also discuss the optimization of their attachment to carrier protein CRM197, affording antigenic and immunogenic conjugates ready for animal vaccination...
October 6, 2016: Tetrahedron
https://www.readbyqxmd.com/read/28187299/development-of-biohybrid-immuno-selective-membranes-for-target-antigen-recognition
#15
Francesca Militano, Teresa Poerio, Rosalinda Mazzei, Simona Salerno, Loredana De Bartolo, Lidietta Giorno
Membranes are gaining increasing interest in solid-phase analytical assay and biosensors applications, in particular as functional surface for bioreceptors immobilization and stabilization as well as for the concentration of target molecules in microsystems. In this work, regenerated cellulose immuno-affinity membranes were developed and they were used for the selective capture of interleukin-6 (IL-6) as targeted antigen. Protein G was covalently linked on the membrane surface and it was successfully used for the oriented site-specific antibody immobilization...
February 3, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28184223/fc-engineering-for-developing-therapeutic-bispecific-antibodies-and-novel-scaffolds
#16
REVIEW
Hongyan Liu, Abhishek Saxena, Sachdev S Sidhu, Donghui Wu
Therapeutic monoclonal antibodies have become molecules of choice to treat autoimmune disorders, inflammatory diseases, and cancer. Moreover, bispecific/multispecific antibodies that target more than one antigen or epitope on a target cell or recruit effector cells (T cell, natural killer cell, or macrophage cell) toward target cells have shown great potential to maximize the benefits of antibody therapy. In the past decade, many novel concepts to generate bispecific and multispecific antibodies have evolved successfully into a range of formats from full bispecific immunoglobulin gammas to antibody fragments...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28183369/identification-of-potential-protein-partners-that-bind-to-the-variant-surface-glycoprotein-in-trypanosoma-equiperdum
#17
Liomary M Carrasquel, José L Escalona, Alvaro Acosta-Serrano, Yurong Guo, José Bubis
Trypanosoma equiperdum possesses a dense coat of a variant surface glycoprotein (VSG) that is used to evade the host immune response by a process known as antigenic variation. Soluble and membrane forms of the predominant VSG from the Venezuelan T. equiperdum TeAp-N/D1 strain (sVSG and mVSG, respectively) were purified to homogeneity; and antibodies against sVSG and mVSG were raised, isolated, and employed to produce anti-idiotypic antibodies that structurally mimic the VSG surface. Prospective VSG-binding partners were initially detected by far-Western blots, and then by immunoblots using the generated anti-idiotypic antibodies...
February 10, 2017: Parasitology
https://www.readbyqxmd.com/read/28183062/thioredoxin-trxo1-interacts-with-proliferating-cell-nuclear-antigen-pcna-and-its-overexpression-affects-the-growth-of-tobacco-cell-culture
#18
Aingeru Calderón, Ana Ortiz-Espín, Raquel Iglesias-Fernández, Pilar Carbonero, Federico Vicente Pallardó, Francisca Sevilla, Ana Jiménez
Thioredoxins (Trxs), key components of cellular redox regulation, act by controlling the redox status of many target proteins, and have been shown to play an essential role in cell survival and growth. The presence of a Trx system in the nucleus has received little attention in plants, and the nuclear targets of plant Trxs have not been conclusively identified. Thus, very little is known about the function of Trxs in this cellular compartment. Previously, we studied the intracellular localization of PsTrxo1 and confirmed its presence in mitochondria and, interestingly, in the nucleus under standard growth conditions...
January 31, 2017: Redox Biology
https://www.readbyqxmd.com/read/28177783/expression-purification-and-renaturation-of-recombinant-peptide-based-hiv-vaccine-in-escherichia-coli
#19
Wei He, Jiayi Shu, Jian Zhang, Zhihua Liu, Jianqing Xu, Xia Jin, Xuedong Wang
One of the new HIV vaccine strategies is to design epitope-based immunogen. We previously predicted 20 potential HIV epitopes using bioinformatics approaches. The combination of these 20 epitopes has a theoretical coverage of 98.1% of the population for both the prevalent HIV genotypes and Chinese human leukocyte antigen (HLA) DR types. In order to test whether this vaccine is immunogenic in vivo, a corresponding antigen needs to be prepared. To this end, we constructed recombinant plasmid containing DNA encoding 300 amino acids of this vaccine protein, and then transformed Escherichia coli BL21 (DE3) cells with the plasmid...
January 27, 2017: Canadian Journal of Microbiology
https://www.readbyqxmd.com/read/28177609/fine-epitope-mapping-of-two-antibodies-neutralizing-the-bordetella-adenylate-cyclase-toxin
#20
Xianzhe Wang, James A Stapleton, Justin R Klesmith, Erik L Hewlett, Tim A Whitehead, Jennifer A Maynard
Adenylate cyclase toxin (ACT) is an important B. pertussis virulence factor that is not included in current acellular pertussis vaccines. We previously demonstrated that immunization with the repeat-in-toxin (RTX) domain of ACT elicits neutralizing antibodies in mice and discovered the first two antibodies to neutralize ACT activities by occluding the receptor-binding site. Here, we fully characterized these antibodies and their epitopes. Both antibodies bind ACT with low nanomolar affinity and cross-react with ACT homologs produced by B...
February 8, 2017: Biochemistry
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