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Antibody purification

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https://www.readbyqxmd.com/read/28229045/occurrence-of-giardia-cryptosporidium-and-entamoeba-in-wild-rhesus%C3%A2-macaques-macaca-mulatta-living-in-urban-and-semi-rural-north-west-india
#1
John J Debenham, Kristoffer Tysnes, Sandhya Khunger, Lucy J Robertson
Giardia duodenalis, Cryptosporidium spp., and Entamoeba spp. are intestinal protozoa capable of infecting a range of host species, and are important causes of human morbidity and mortality. Understanding their epidemiology is important, both for public health and for the health of the animals they infect. This study investigated the occurrence of these protozoans in rhesus macaques (Macaca mulatta) in India, with the aim of providing preliminary information on the potential for transmission of these pathogens between macaques and humans...
April 2017: International Journal for Parasitology. Parasites and Wildlife
https://www.readbyqxmd.com/read/28224978/methylated-antibody-affinity-purification-to-improve-proteomic-identification-of-plant-rna-polymerase-pol-v-complex-and-the-interacting-proteins
#2
Guochen Qin, Jun Ma, Xiaomei Chen, Zhaoqing Chu, Yi-Min She
Affinity purification followed by enzymatic digestion and mass spectrometry has been widely utilized for the sensitive detection of interacting proteins and protein complexes in various organisms. In plants, the method is technically challenging due to the low abundance proteins, non-specific binding and difficulties of eluting interacting proteins from antibody beads. In this report, we describe a strategy to modify antibodies by reductive methylation of lysines without affecting their binding properties, followed by on-bead digestion of bound proteins with endoproteinase Lys-C...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28224267/selection-of-dna-aptamers-for-extra-cellular-domain-of-human-epidermal-growth-factor-receptor-2-to-detect-her2-positive-carcinomas
#3
A Sett, B B Borthakur, U Bora
BACKGROUND: Human epidermal growth factor receptor 2 (Her2, an orphan receptor of ErbB family) is considered as an important biomarker as it plays a key role in the development and progression of aggressive types of breast, ovarian, stomach and gastric cancer. In the present study, we developed novel DNA aptamers against the extra-cellular domain (ECD) of Her2 protein for detection of Her2-positive carcinomas. METHODS: We cloned and expressed Her2-ECD protein in E...
February 21, 2017: Clinical & Translational Oncology
https://www.readbyqxmd.com/read/28222997/integrated-analysis-of-recombinant-bpv-1-l1-protein-for-the-production-of-a-bovine-papillomavirus-vlp-vaccine
#4
Diego Grando Módolo, Rodrigo Pinheiro Araldi, Jacqueline Mazzuchelli-de-Souza, Alexandre Pereira, Daniel Carvalho Pimenta, Letícia Maria Zanphorlin, Willy Beçak, Marcelo Menossi, Rita de Cassia Stocco, Rodrigo Franco de Carvalho
Bovine papillomatosis is an infectious disease that is caused by bovine papillomavirus (BPV), which results in important economic losses. However, no BPV vaccines or effective treatment methods are commercially available to date. Moreover, the absence of papillomavirus replication in vitro makes the use of recombinant protein a promising candidate for vaccine formulations. Hence, we developed an integrated study on the L1 capsid protein of BPV-1, obtained from a bacterial expression system, regarding its purification, biosafety, thermostability and immunogenicity...
February 17, 2017: Vaccine
https://www.readbyqxmd.com/read/28222187/a-purified-maa-based-elisa-is-a-useful-tool-for-determining-anti-maa-antibody-titer-with-high-sensitivity
#5
Takasumi Shimomoto, Leonard B Collins, Xianwen Yi, Darcy W Holley, Zhenfa Zhang, Xu Tian, Koji Uchida, Chunguang Wang, Sohvi Hörkkö, Monte S Willis, Avram Gold, Scott J Bultman, Jun Nakamura
Atherosclerosis is widely accepted to be a chronic inflammatory disease, and the immunological response to the accumulation of LDL is believed to play a critical role in the development of this disease. 1,4-Dihydropyridine-type MAA-adducted LDL has been implicated in atherosclerosis. Here, we have demonstrated that pure MAA-modified residues can be chemically conjugated to large proteins without by-product contamination. Using this pure antigen, we established a purified MAA-ELISA, with which a marked increase in anti-MAA antibody titer was determined at a very early stage of atherosclerosis in 3-month ApoE-/- mice fed with a normal diet...
2017: PloS One
https://www.readbyqxmd.com/read/28220811/aptamers-for-respiratory-syncytial-virus-detection
#6
Krisztina Percze, Zoltán Szakács, Éva Scholz, Judit András, Zsuzsanna Szeitner, Corné H van den Kieboom, Gerben Ferwerda, Marien I de Jonge, Róbert E Gyurcsányi, Tamás Mészáros
The identification of the infectious agents is pivotal for appropriate care of patients with viral diseases. Current viral diagnostics rely on selective detection of viral nucleic acid or protein components. In general, detection of proteins rather than nucleic acids is technically more suitable for rapid tests. However, protein-based virus identification methods depend on antibodies limiting the practical applicability of these approaches. Aptamers rival antibodies in target selectivity and binding affinity, and excel in terms of robustness and cost of synthesis...
February 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28220238/application-of-calcium-phosphate-flocculation-in-high-density-cell-culture-fluid-with-high-product-titer-of-monoclonal-antibody
#7
Guoqiang Chen, Zhiguo Su, Feng Li, Hui Fang Liu
The calcium phosphate [Ca3(PO4)2] precipitation was used for improving the clarification efficiency in harvest process of the monoclonal antibody (mAb) containing cell culture fluid (CCF) with high turbidity and product titer. The flocculation conditions (concentration, addition order of flocculants, pH, and operation time), and the effect of flocculants on the mAb physical chemical properties (such as distribution of charge variants and aggregates) and process-related impurities removal (such as DNA and CHOP) were evaluated in this study...
February 20, 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28219449/ex-vivo-18-o-labeling-mass-spectrometry-identifies-a-peripheral-amyloid-%C3%AE-clearance-pathway
#8
Erik Portelius, Niklas Mattsson, Josef Pannee, Henrik Zetterberg, Magnus Gisslén, Hugo Vanderstichele, Eleni Gkanatsiou, Gabriela A N Crespi, Michael W Parker, Luke A Miles, Johan Gobom, Kaj Blennow
BACKGROUND: Proteolytic degradation of amyloid β (Aβ) peptides has been intensely studied due to the central role of Aβ in Alzheimer's disease (AD) pathogenesis. While several enzymes have been shown to degrade Aβ peptides, the main pathway of Aβ degradation in vivo is unknown. Cerebrospinal fluid (CSF) Aβ42 is reduced in AD, reflecting aggregation and deposition in the brain, but low CSF Aβ42 is, for unknown reasons, also found in some inflammatory brain disorders such as bacterial meningitis...
February 20, 2017: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/28218470/maximizing-the-functional-lifetime-of-protein-a-resins
#9
Jennifer Zhang, Sethu Siva, Ryan Caple, Rob Gronke, Sanchayita Ghose
Protein A chromatography is currently the industry gold-standard for monoclonal antibody and Fc-fusion protein purification. The high cost of Protein A however makes resin lifetime and resin reuse an important factor for process economics. Typical resin lifetime studies performed in the industry usually examine the effect of resin re-use on binding capacity, yield, and product quality without answering the fundamental question of what is causing the decrease in performance. A two part mechanistic study was conducted in an attempt to decouple the effect of the two possible factors (resin hydrolysis and/or degradation vs...
February 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28216237/development-of-an-anti-ferret-cd4-monoclonal-antibody-for-the-characterisation-of-ferret-t-lymphocytes
#10
Daniel S Layton, Xiaowen Xiao, John D Bentley, Louis Lu, Cameron R Stewart, Andrew G D Bean, Timothy E Adams
The ferret is an established animal model for a number of human respiratory viral infections, such as influenza virus and more recently, Ebola virus. However, a paucity of immunological reagents has hampered the study of cellular immune responses. Here we describe the development and characterisation of a novel monoclonal antibody (mAb) against the ferret CD4 antigen and the characterisation of ferret CD4 T lymphocytes. Recombinant production and purification of the ferret CD4 ectodomain soluble protein allowed hybridoma generation and the generation of a mAb (FeCD4) showing strong binding to ferret CD4 protein and lymphoid cells by flow cytometry...
February 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28214589/enhanced-expression-and-purification-of-camelid-single-domain-vhh-antibodies-from-classical-inclusion-bodies
#11
Maristella Maggi, Claudia Scotti
Single domain antibodies (sdAbs) are small antigen-binding domains derived from naturally occurring, heavy chain-only immunoglobulins isolated from camelid and sharks. They maintain the same binding capability of full-length IgGs but with improved thermal stability and permeability, which justifies their scientific, medical and industrial interest. Several described recombinant forms of sdAbs have been produced in different hosts and with different strategies. Here we present an optimized method for a time-saving, high yield production and extraction of a poly-histidine-tagged sdAb from Escherichia coli classical inclusion bodies...
February 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28212428/a-novel-hemagglutinin-protein-produced-in-bacteria-protects-chickens-against-h5n1-highly-pathogenic-avian-influenza-viruses-by-inducing-h5-subtype-specific-neutralizing-antibodies
#12
Violetta Sączyńska, Agnieszka Romanik, Katarzyna Florys, Violetta Cecuda-Adamczewska, Małgorzata Kęsik-Brodacka, Krzysztof Śmietanka, Monika Olszewska, Katarzyna Domańska-Blicharz, Zenon Minta, Bogusław Szewczyk, Grażyna Płucienniczak, Andrzej Płucienniczak
The highly pathogenic (HP) H5N1 avian influenza viruses (AIVs) cause a mortality rate of up to 100% in infected chickens and pose a permanent pandemic threat. Attempts to obtain effective vaccines against H5N1 HPAIVs have focused on hemagglutinin (HA), an immunodominant viral antigen capable of eliciting neutralizing antibodies. The vast majority of vaccine projects have been performed using eukaryotic expression systems. In contrast, we used a bacterial expression system to produce vaccine HA protein (bacterial HA) according to our own design...
2017: PloS One
https://www.readbyqxmd.com/read/28209207/recombinant-%C3%AE-actinin-subunit-antigens-of-trichomonas-vaginalis-as-potential-vaccine-candidates-in-protecting-against-trichomoniasis
#13
Yi-Ting Xie, Jiang-Mei Gao, Ya-Ping Wu, Petrus Tang, Geoff Hide, De-Hua Lai, Zhao-Rong Lun
BACKGROUND: Human trichomoniasis caused by Trichomonas vaginalis is one of the most common sexually transmitted diseases with more than 200 million cases worldwide. It has caused a series of health problems to patients. For prevention and control of infectious diseases, vaccines are usually considered as one of the most cost-efficient tools. However, until now, work on the development of T. vaginalis vaccines is still mainly focused on the screening of potential immunogens. Alpha-actinin characterized by high immunogenicity in T...
February 16, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28202756/a-glycosylation-benchmark-profile-for-hiv-1-envelope-glycoprotein-production-based-on-eleven-env-trimers
#14
Eden P Go, Haitao Ding, Shijian Zhang, Rajesh P Ringe, Nathan Nicely, David Hua, Robert T Steinbock, Michael Golabek, James Alin, S Munir Alam, Albert Cupo, Barton F Haynes, John C Kappes, John P Moore, Joseph G Sodroski, Heather Desaire
HIV-1 envelope glycoprotein (Env) glycosylation is important because individual glycans are components of multiple broadly neutralizing antibody epitopes, while shielding other sites that might otherwise be immunogenic. The glycosylation on Env is influenced by a variety of factors, including the genotype of the protein, the cell line used for its expression, and the details of the construct design. Here, we used a mass spectrometry-based approach to map the complete glycosylation profile at every site in multiple HIV-1 Env trimers, accomplishing two goals: 1) We determined which glycosylation sites contain conserved glycan profiles across many trimeric Envs...
February 15, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28193502/development-of-a-preclinical-211-rn-211-at-generator-system-for-targeted-alpha-therapy-research-with-211-at
#15
Jason R Crawford, Hua Yang, Peter Kunz, D Scott Wilbur, Paul Schaffer, Thomas J Ruth
INTRODUCTION: The availability of (211)At for targeted alpha therapy research can be increased by the (211)Rn/(211)At generator system, whereby (211)At is produced by (211)Rn electron capture decay. This study demonstrated the feasibility of using generator-produced (211)At to label monoclonal antibody (BC8, anti-human CD45) for preclinical use, following isolation from the (207)Po contamination also produced by these generators (by (211)Rn α-decay). METHODS: (211)Rn was produced by (211)Fr electron capture decay following mass separated ion beam implantation and chemically isolated in liquid alkane hydrocarbon (dodecane)...
January 29, 2017: Nuclear Medicine and Biology
https://www.readbyqxmd.com/read/28193052/characterization-of-chain-pairing-variants-of-bispecific-igg-expressed-in-a-single-host-cell-by-high-resolution-native-and-denaturing-mass-spectrometry
#16
Luis Schachner, Guanghui Han, Michael Dillon, Jianhui Zhou, Luke McCarty, Diego Ellerman, Yiyuan Yin, Christoph Spiess, Jennie R Lill, Paul J Carter, Wendy Sandoval
Bispecific antibodies, including bispecific IgG, show some promise in clinical trials as a means to extend the therapeutic potential of antibodies. Bispecific IgG can be made by separate expression and purification of each parent half antibody followed by in vitro reconstitution. Generating bispecific IgG by coexpression of two different light and heavy chains in a single host cell is potentially more efficient because it obviates the need for two separate cell lines and purification processes. However, this workflow may produce unwanted mispaired IgG species in addition to the desired bispecific IgG...
December 20, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/28186329/effects-of-antibody-disulfide-bond-reduction-on-purification-process-performance-and-final-drug-substance-stability
#17
Wai Keen Chung, Brian Russell, Yanhong Yang, Michael Handlogten, Suzanne Hudak, Mingyan Cao, Jihong Wang, David Robbins, Sanjeev Ahuja, Min Zhu
Antibody disulfide bond reduction during monoclonal antibody (mAb) production is a phenomenon that has been attributed to the reducing enzymes from CHO cells acting on the mAb during the harvest process. However, the impact of antibody reduction on the downstream purification process has not been studied. During the production of an IgG2 mAb, antibody reduction was observed in the harvested cell culture fluid (HCCF), resulting in high fragment levels. In addition, aggregate levels increased during the low pH treatment step in the purification process...
February 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28183045/new-process-for-purifying-high-purity-%C3%AE-1-antitrypsin-from-cohn-fraction-iv-by-chromatography-a-promising-method-for-the-better-utilization-of-plasma
#18
Chaoji Huangfu, Jinchao Zhang, Yuyuan Ma, Junting Jia, Maomin Lv, Xiong Zhao, Jingang Zhang
α1-antitrypsin (AAT) is a 52kDa serine protease inhibitor that is abundant in plasma. It is synthesized mainly by hepatic cells, and widely used to treat patients with emphysema due to congenital deficiency of AAT. A new isolation method for the purification of AAT from Cohn Fraction IV (Cohn F IV) is described. Cohn F IV is usually discarded as a byproduct from Cohn process. Using Cohn F IV as starting material does not interfere with the production of other plasma proteins and the cost of purification could be reduced greatly...
February 1, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28181316/an-improved-method-for-purification-and-refolding-of-recombinant-hiv-vif-expressed-in-e-coli
#19
Carlos A Duarte, Mickel Palomino
Virion infectivity factor (Vif) is a 23 KDa protein which protects HIV-1 from deamination of its proviral DNA by APOBEC3G. The active form of Vif is a multimer that interacts simultaneously with CBF-beta, the Elongin B and C subunits, Cullin 5 and APOBEC3G to form a ubiquitin ligase complex targeting the latter for degradation. Vif clearly represents an attractive target for developing novel antiviral drugs for the therapy of HIV/AIDS, and this goal requires a source of well folded, readily available protein...
February 8, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28178596/automated-2d-hplc-method-for-characterization-of-protein-aggregation-with-in-line-fraction-collection-device
#20
Abasha Williams, Erik K Read, Cyrus D Agarabi, Scott Lute, Kurt A Brorson
Monoclonal antibodies are mainly produced by mammalian cell culture, which due to its complexity, results in a wide range of product variants/isoforms. With the growing implementation of Quality by Design (QbD) and Process Analytical Technology (PAT) in drug manufacturing, monitoring and controlling quality attributes within a predefined range during manufacturing may provide added consistency to product quality. To implement these concepts, more robust analytical tools could reduce the time needed for monitoring quality attributes during upstream processing...
January 25, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
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