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Antibody purification

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https://www.readbyqxmd.com/read/28822126/dimethyl-labeling-based-quantification-of-the-lysine-acetylome-and-proteome-of-plants
#1
Ines Lassowskat, Markus Hartl, Fabian Hosp, Paul J Boersema, Matthias Mann, Iris Finkemeier
Photorespiratory enzymes in different cellular compartments have been reported to be posttranslational modified by phosphorylation, disulfide formation, S-nitrosylation, glutathionylation, and lysine acetylation. However, not much is known yet about the function of these modifications to regulate the activities, localizations, or interactions of the proteins in this metabolic pathway. Hence, it will be of great importance to study these modifications and their temporal and conditional occurrence in more detail...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28807549/a-multiplexed-microfluidic-toolbox-for-the-rapid-optimization-of-affinity-driven-partition-in-aqueous-two-phase-systems
#2
Eduardo J S Bras, Ruben R G Soares, Ana M Azevedo, Pedro Fernandes, Miguel Arévalo-Rodríguez, Virginia Chu, João P Conde, M Raquel Aires-Barros
Antibodies and other protein products such as interferons and cytokines are biopharmaceuticals of critical importance which, in order to be safely administered, have to be thoroughly purified in a cost effective and efficient manner. The use of aqueous two-phase extraction (ATPE) is a viable option for this purification, but these systems are difficult to model and optimization procedures require lengthy and expensive screening processes. Here, a methodology for the rapid screening of antibody extraction conditions using a microfluidic channel-based toolbox is presented...
August 5, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28807316/vaccination-with-eimeria-tenella-elongation-factor-1%C3%AE-recombinant-protein-induces-protective-immunity-against-e-tenella-and-e-maxima-infections
#3
Rui-Qing Lin, Hyun S Lillehoj, Seung Kyoo Lee, Sungtaek Oh, Alfredo Panebra, Erik P Lillehoj
Avian coccidiosis is caused by multiple species of the apicomplexan protozoan, Eimeria, and is one of the most economically devastating enteric diseases for the poultry industry worldwide. Host immunity to Eimeria infection, however, is relatively species-specific. The ability to immunize chickens against different species of Eimeria using a single vaccine will have a major beneficial impact on commercial poultry production. In this paper, we describe the molecular cloning, purification, and vaccination efficacy of a novel Eimeria vaccine candidate, elongation factor-1α (EF-1α)...
August 30, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28806153/expression-purification-and-refolding-of-human-lipocalin-6-and-production-of-a-monoclonal-antibody-against-this-protein
#4
Jiong Chen, Wei Feng, Yue Zhao, Yaqin Li, Fei Zhan
Human lipocalin 6 (hLCN6) is a member of the lipocalin family, which is a group of structurally conserved hydrophobic ligand binding proteins, and widely distributed in animal, plant, and bacteria. Specific expression of hLCN6 in the epididymis and localization of this protein on the surface of spermatozoa suggest a role played by hLCN6, which may function as a transporter to carry ligands in the epididymal channel. However, the role of hLCN6 in sperm maturation has been largely unknown due to the lack of effective antibodies...
August 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28805498/gingiskhan%C3%A2-protease-cleavage-allows-a-high-throughput-antibody-to-fab-conversion-enabling-direct-functional-assessment-during-lead-identification-of-human-monoclonal-and-bispecific-igg1-antibodies
#5
Jörg Moelleken, Manuel Endesfelder, Christian Gassner, Sabine Lingke, Simone Tomaschek, Oksana Tyshchuk, Stefan Lorenz, Ulrike Reiff, Michael Mølhøj
The determination of the binding strength of immunoglobulins (IgGs) to targets can be influenced by avidity when the targets are soluble di- or multimeric proteins, or associated to cell surfaces, including surfaces introduced from heterogeneous assays. However, for the understanding of the contribution of a second drug-to-target binding site in molecular design, or for ranking of monovalent binders during lead identification, affinity-based assessment of the binding strength is required. Typically, monovalent binders like antigen-binding fragments (Fabs) are generated by proteolytic cleavage with papain, which often results in a combination of under- and over-digestion, and requires specific optimization and chromatographic purification of the desired Fabs...
August 14, 2017: MAbs
https://www.readbyqxmd.com/read/28803008/two-step-large-volume-magnetic-separation-combined-with-pcr-assay-for-sensitive-detection-of-listeria-monocytogenes-in-pasteurized-milk
#6
Dan Luo, Xiaolin Huang, Yan Mao, Chaochao Chen, Fulai Li, Hengyi Xu, Yonghua Xiong
Immunomagnetic separation (IMS) is an effective tool for the preconcentration and purification of food-borne pathogens from complex food samples because of its high capture efficiency (CE). In conventional IMS, antibodies are usually conjugated on the surface of magnetic beads (MB); the random orientation and conformation changes of antibodies on the MB surface can decrease their bioactivity. Moreover, the Brownian motion of immobilized antibodies is weakened, thereby rendering their binding efficiency with bacteria lower than that of free antibodies...
August 9, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28802994/expression-and-purification-of-norovirus-virus-like-particles-in-escherichia-coli-and-their-immunogenicity-in-mice
#7
Yuqi Huo, Xin Wan, Tong Ling, Jie Wu, Wenhui Wang, Shuo Shen
Norovirus (NoV) virus like particles (VLPs) produced in Spodoptera frugiperda (Sf9) cells have been tested in human volunteers as vaccine candidate and were shown to be protective against NoV induced acute gastroenteritis. In this study, prevailing Sydney-2012-like NoV major capsid protein gene with or without N-terminal deletions (N26 and N38, 26 and 38 amino acids deleted from N terminus,respectively) were sub-cloned into prokaryotic expression vector, pCold III and pCold IV. Soluble and insoluble proteins were detected for both vectors after induction and higher levels of protein expression were observed for constructs pCold III-N26 and pCold III-N38...
August 9, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28797234/succinyl-proteome-profiling-of-dendrobium-officinale-an-important-traditional-chinese-orchid-herb-revealed-involvement-of-succinylation-in-the-glycolysis-pathway
#8
Shangguo Feng, Kaili Jiao, Hong Guo, Mengyi Jiang, Juan Hao, Huizhong Wang, Chenjia Shen
BACKGROUND: Lysine succinylation is a ubiquitous and important protein post-translational modification in various eukaryotic and prokaryotic cells. However, its functions in Dendrobium officinale, an important traditional Chinese orchid herb with high polysaccharide contents, are largely unknown. RESULTS: In our study, LC-MS/MS was used to identify the peptides that were enriched by immune-purification with a high-efficiency succinyl-lysine antibody. In total, 314 lysine succinylation sites in 207 proteins were identified...
August 10, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28795201/downstream-processing-technologies-capturing-and-final-purification-opportunities-for-innovation-change-and-improvement-a-review-of-downstream-processing-developments-in-protein-purification
#9
Nripen Singh, Sibylle Herzer
Increased pressure on upstream processes to maximize productivity has been crowned with great success, although at the cost of shifting the bottleneck to purification. As drivers were economical, focus is on now on debottlenecking downstream processes as the main drivers of high manufacturing cost. Devising a holistically efficient and economical process remains a key challenge. Traditional and emerging protein purification strategies with particular emphasis on methodologies implemented for the production of recombinant proteins of biopharmaceutical importance are reviewed...
August 10, 2017: Advances in Biochemical Engineering/biotechnology
https://www.readbyqxmd.com/read/28795022/the-potency-of-chicken-egg-yolk-immunoglobulin-igy-specific-as-immunotherapy-to-mycobacterium-tuberculosis-infection
#10
Sri Agus Sudjarwo, Koerniasari Eraiko, Giftania Wardani Sudjarwo, Koerniasari
The aim of this study was to characterize of chicken egg yolk immunoglobulins (IgYs) specific as immunotherapy to Mycobacterium tuberculosis complex (MTBC) infection. Lohmann laying hens were immunized intramuscularly with antigenic of MTBC. Egg yolk was separated from egg white, and IgY antibody was then purified by multiple polyethylene glycols 6000 extraction and ammonium sulfate purification steps. The IgY anti-MTBC concentration in egg yolk increased at 2 weeks and it reached a maximum at 4 weeks after immunization...
July 2017: Journal of Advanced Pharmaceutical Technology & Research
https://www.readbyqxmd.com/read/28783371/molecular-characterization-and-biological-activity-of-interferon-%C3%AE-in-indian-peafowl-pavo-cristatus
#11
Hongjing Zhao, Yu Wang, Juanjuan Liu, Yizhi Shao, Jinglun Li, Hongliang Chai, Mingwei Xing
Interferon-α (IFN-α) genes have been cloned from a variety of animals, but information about Indian peafowl Pavo cristatus IFN-α (PcIFN-α) has not been reported. In this study, a PcIFN-α gene was amplified, which encoded a protein of 193 amino acids with a 26-amino acid signal peptide sharing 72.16-95.70% identity with other avians in Aves. After expression in prokaryote, purification, and renaturation, PcIFN-α was analyzed for its physicochemical property and antiviral activity. Cytopathic inhibition test showed that the recombinant PcIFN-α could inhibit the replication of vesicular stomatitis virus (VSV), Newcastle disease virus (NDV), and avian influenza virus (AIV) on chicken embryo fibroblasts...
August 7, 2017: DNA and Cell Biology
https://www.readbyqxmd.com/read/28774114/fc-binding-ligands-of-immunoglobulin-g-an-overview-of-high-affinity-proteins-and-peptides
#12
REVIEW
Weonu Choe, Trishaladevi A Durgannavar, Sang J Chung
The rapidly increasing application of antibodies has inspired the development of several novel methods to isolate and target antibodies using smart biomaterials that mimic the binding of Fc-receptors to antibodies. The Fc-binding domain of antibodies is the primary binding site for e.g., effector proteins and secondary antibodies, whereas antigens bind to the Fab region. Protein A, G, and L, surface proteins expressed by pathogenic bacteria, are well known to bind immunoglobulin and have been widely exploited in antibody purification strategies...
December 8, 2016: Materials
https://www.readbyqxmd.com/read/28771492/production-of-an-anti-a%C3%AE-antibody-fragment-in-pichia-pastoris-and-in-vitro-and-in-vivo-validation-of-its-therapeutic-effect
#13
Laia Montoliu-Gaya, Gisela Esquerda-Canals, Silvia Bronsoms, Sandra Villegas
ScFv-h3D6 has been shown as an efficient therapy in the 3xTg-AD mouse model of Alzheimer's Disease. Because one of the major bottlenecks for the therapeutic uses of proteins produced in Escherichia coli is their potential contamination with endotoxins, LPS were extensively removed by a rather low-efficient, expensive, and time-consuming purification step. In addition, disulfide scrambling is favored in the reducing bacterial cytoplasm albeit the use of reductase deficient strains. To overcome these hurdles, as well as to improve the yield, the yeast Pichia pastoris, an endotoxin-free host system for recombinant protein production, has been used to produce scFv-h3D6, both in flask and in a fed-batch bioreactor...
2017: PloS One
https://www.readbyqxmd.com/read/28771197/the-susceptibilities-of-respiratory-syncytial-virus-to-nucleolin-receptor-blocking-and-antibody-neutralization-are-dependent-upon-the-method-of-virus-purification
#14
Leanne M Bilawchuk, Cameron D Griffiths, Lionel D Jensen, Farah Elawar, David J Marchant
Respiratory Syncytial Virus (RSV) that is propagated in cell culture is purified from cellular contaminants that can confound experimental results. A number of different purification methods have been described, including methods that utilize fast protein liquid chromatography (FPLC) and gradient ultracentrifugation. Thus, the constituents and experimental responses of RSV stocks purified by ultracentrifugation in sucrose and by FPLC were analyzed and compared by infectivity assay, Coomassie stain, Western blot, mass spectrometry, immuno-transmission electron microscopy (TEM), and ImageStream flow cytometry...
August 3, 2017: Viruses
https://www.readbyqxmd.com/read/28765476/annexin-a11-is-targeted-by-igg4-and-igg1-autoantibodies-in-igg4-related-disease
#15
Lowiek M Hubers, Harmjan Vos, Alex R Schuurman, Robin Erken, Ronald P Oude Elferink, Boudewijn Burgering, Stan F J van de Graaf, Ulrich Beuers
OBJECTIVE: Immunoglobulin G4-related disease (IgG4-RD) is a multiorgan immune-mediated disease that predominantly affects the biliary tract (IgG4-associated cholangitis, IAC) and pancreas (autoimmune pancreatitis, AIP). We recently identified highly expanded IgG4+ B-cell receptor clones in blood and affected tissues of patients with IAC/AIP suggestive of specific (auto)antigenic stimuli involved in initiating and/or maintaining the inflammatory response. This study aimed to identify (auto)antigen(s) that are responsible for the clonal expansion of IgG4+ B cells in IgG4-RD...
August 1, 2017: Gut
https://www.readbyqxmd.com/read/28764709/antibody-responses-to-two-new-lactococcus-lactis-produced-recombinant-pfs48-45-and-pfs230-proteins-increase-with-age-in-malaria-patients-living-in-the-central-region-of-ghana
#16
Festus K Acquah, Evans K Obboh, Kwame Asare, Johnson N Boampong, Samuel Victor Nuvor, Susheel K Singh, Michael Theisen, Kim C Williamson, Linda Eva Amoah
BACKGROUND: Recent advances in malaria control efforts have led to an increased number of national malaria control programmes implementing pre-elimination measures and demonstrated the need to develop new tools to track and control malaria transmission. Key to understanding transmission is monitoring the prevalence and immune response against the sexual stages of the parasite, known as gametocytes, which are responsible for transmission. Sexual-stage specific antigens, Pfs230 and Pfs48/45, have been identified and shown to be targets for transmission blocking antibodies, but they have been difficult to produce recombinantly in the absence of a fusion partner...
August 1, 2017: Malaria Journal
https://www.readbyqxmd.com/read/28762517/immunoaffinity-based-methods-are-superior-to-kits-for-purification-of-prostate-derived-extracellular-vesicles-from-plasma-samples
#17
Sabine I Brett, Fabrice Lucien, Charles Guo, Karla C Williams, Yohan Kim, Paul N Durfee, C J Brinker, Joseph I Chin, Jun Yang, Hon S Leong
BACKGROUND: The ability to isolate extracellular vesicles (EVs) such as exosomes or microparticles is an important method that is currently not standardized. While commercially available kits offer purification of EVs from biofluids, such purified EV samples will also contain non-EV entities such as soluble protein and nucleic acids that could confound subsequent experimentation. Ideally, only EVs would be isolated and no soluble protein would be present in the final EV preparation. METHODS: We compared commercially available EV isolation kits with immunoaffinity purification techniques and evaluated our final EV preparations using atomic force microscopy (AFM) and nanoscale flow cytometry (NFC)...
August 1, 2017: Prostate
https://www.readbyqxmd.com/read/28758805/immunoaffinity-techniques-coupled-to-mass-spectrometry-for-the-analysis-of-human-peptide-hormones-advances-and-applications
#18
Andreas Thomas, Wilhelm Schänzer, Mario Thevis
The accurate and comprehensive determination of peptide hormones from biological fluids has represented a considerable challenge to analytical chemists for decades. Besides long-established bioanalytical ligand binding assays (or ELISA, RIA, etc.), more and more mass spectrometry-based methods have been developed recently for purposes commonly referred to as targeted proteomics. Eventually the combination of both, analyte extraction by immunoaffinity and subsequent detection by mass spectrometry, has shown to synergistically enhance the test methods' performance characteristics...
July 31, 2017: Expert Review of Proteomics
https://www.readbyqxmd.com/read/28757080/a-fusion-receptor-as-a-safety-switch-detection-and-purification-biomarker-for-adoptive-transferred-t-cells
#19
Xiuqi Wu, Bizhi Shi, Jiqin Zhang, Zhimin Shi, Shengmeng Di, Minliang Fan, Huiping Gao, Hai Wang, Jianren Gu, Hua Jiang, Zonghai Li
The incorporation of an endogenous safety switch represents a rational strategy for the control of toxicities following the administration of adoptive T cell therapies. An ideal safety switch should be capable of depleting the transferred T cells with minimal injury to normal tissues. We generated a fusion receptor by engineering a cryptic 806 epitope of human epidermal growth factor receptor (EGFR) into the N terminus of the full-length human folate receptor 1 (FOLR1), designated as FR806. The expression of FR806 allows transduced T cells to be targeted with CH12, a monoclonal antibody recognizing the 806 epitope, but not wild-type EGFR in healthy tissues...
July 3, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28757031/genetic-characteristics-pathogenicity-and-immunogenicity-associated-with-cell-adaptation-of-a-virulent-genotype-2b-porcine-epidemic-diarrhea-virus
#20
Sunhee Lee, Kyu-Yeol Son, Yun-Hee Noh, Seung-Chul Lee, Hwan-Won Choi, In-Joong Yoon, Changhee Lee
Porcine epidemic diarrhea virus (PEDV) has emerged or re-emerged worldwide, posing a significant financial threat to major pig-producing countries. In the present study, a virulent Korean pandemic PEDV strain, KNU-141112, was serially propagated in Vero cells for up to 100 passages. Through cell culture adaptation, we obtained four distinct deletion (DEL) mutants by plaque purification followed by nucleotide sequencing of the spike (S)/ORF3 gene-coding region, which were designated KNU-141112-S DEL2, -S DEL5, -S DEL2/ORF3, and -S DEL5/ORF3...
August 2017: Veterinary Microbiology
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