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Recombinant antibody

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https://www.readbyqxmd.com/read/29150665/a-streamlined-cloning-workflow-minimising-the-time-to-strain-pipeline-for-pichia-pastoris
#1
Kate E Royle, Karen Polizzi
Although recent advances in E. coli self-assembly have greatly simplified cloning, these have not yet been harnessed for the high-throughput generation of expression strains in the early research and discovery phases of biopharmaceutical production. Here, we have refined the technique and incorporated it into a streamlined workflow for the generation of Pichia pastoris expression strains, reducing the timeline by a third and removing the reliance on DNA editing enzymes, which often require troubleshooting and increase costs...
November 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29149197/sieve-analysis-of-breakthrough-hiv-1-sequences-in-hvtn-505-identifies-vaccine-pressure-targeting-the-cd4-binding-site-of-env-gp120
#2
Allan C deCamp, Morgane Rolland, Paul T Edlefsen, Eric Sanders-Buell, Breana Hall, Craig A Magaret, Andrew J Fiore-Gartland, Michal Juraska, Lindsay N Carpp, Shelly T Karuna, Meera Bose, Steven LePore, Shana Miller, Annemarie O'Sullivan, Kultida Poltavee, Hongjun Bai, Kalpana Dommaraju, Hong Zhao, Kim Wong, Lennie Chen, Hasan Ahmed, Derrick Goodman, Matthew Z Tay, Raphael Gottardo, Richard A Koup, Robert Bailer, John R Mascola, Barney S Graham, Mario Roederer, Robert J O'Connell, Nelson L Michael, Merlin L Robb, Elizabeth Adams, Patricia D'Souza, James Kublin, Lawrence Corey, Daniel E Geraghty, Nicole Frahm, Georgia D Tomaras, M Juliana McElrath, Lisa Frenkel, Sheila Styrchak, Sodsai Tovanabutra, Magdalena E Sobieszczyk, Scott M Hammer, Jerome H Kim, James I Mullins, Peter B Gilbert
Although the HVTN 505 DNA/recombinant adenovirus type 5 vector HIV-1 vaccine trial showed no overall efficacy, analysis of breakthrough HIV-1 sequences in participants can help determine whether vaccine-induced immune responses impacted viruses that caused infection. We analyzed 480 HIV-1 genomes sampled from 27 vaccine and 20 placebo recipients and found that intra-host HIV-1 diversity was significantly lower in vaccine recipients (P ≤ 0.04, Q-values ≤ 0.09) in Gag, Pol, Vif and envelope glycoprotein gp120 (Env-gp120)...
2017: PloS One
https://www.readbyqxmd.com/read/29146448/sialic-acid-binding-lectins-sabls-from-solen-grandis-function-as-prrs-ensuring-immune-recognition-and-bacterial-clearance
#3
Xiumei Wei, Dinglong Yang, Huiying Li, Hailin Jiang, Xiangquan Liu, Qin Zhang, Jialong Yang
Sialic acid-binding lectins (SABLs) are ubiquitous ancient molecules with binding properties to N-acetyl or N-glycolyl carbohydrates, and play crucial roles in both adaptive and innate immune responses. In present study, recombinant protein and antibodies of two SABLs from mollusk Solen grandis (SgSABL-1 and SgSABL-2) were prepared to investigate their functions in innate immunity. The recombinant protein of SgSABL-1 (rSgSABL-1) could bind LPS, PGN and β-glucan in vitro, while rSgSABL-2 could only bind PGN rather than LPS and β-glucan...
November 13, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29146379/lt-adjuvant-modulates-epitope-specificity-and-improves-the-efficacy-of-murine-antibodies-elicited-by-sublingual-vaccination-with-the-n-terminal-domain-of-streptococcus-mutans-p1
#4
Milene Tavares Batista, Ewerton Lucena Ferreira, Gisela de Souza Pereira, Phillip Stafford, Denicar Lina Nascimento Fabris Maeda, Juliana Falcão Rodrigues, L Jeannine Brady, Stephen Albert Johnston, Luís Carlos de Souza Ferreira, Rita de Cássia Café Ferreira
In this study, we evaluated the immunogenicity, protective efficacy and peptide-based immune signatures of antibodies raised in mice after sublingual immunization with a recombinant form of the P1 (aka AgI/II, PAc) adhesin (P139-512) of Streptococcus mutans, a major etiological agent of dental caries. Sublingual administration of P139-512 in combination with the mucosal adjuvant LTK4R (a derivative of heat-labile LT toxin) induced strong and long-lasting systemic and mucosal immune responses. Incorporation of the adjuvant resulted in an enhancement of the anti-adhesive and anti-colonization activity against S...
November 13, 2017: Vaccine
https://www.readbyqxmd.com/read/29146152/improved-antifungal-activity-of-barley-derived-chitinase-i-gene-that-overexpress-a-32kda-recombinant-chitinase-in-escherichia-coli-host
#5
Nida Toufiq, Bushra Tabassum, Muhammad Umar Bhatti, Anwar Khan, Muhammad Tariq, Naila Shahid, Idrees Ahmad Nasir, Tayyab Husnain
Agricultural crops suffer many diseases, including fungal and bacterial infections, causing significant yield losses. The identification and characterisation of pathogenesis-related protein genes, such as chitinases, can lead to reduction in pathogen growth, thereby increasing tolerance against fungal pathogens. In the present study, the chitinase I gene was isolated from the genomic DNA of Barley (Hordeum vulgare L.) cultivar, Haider-93. The isolated DNA was used as template for the amplification of the ∼935bp full-length chitinase I gene...
October 31, 2017: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/29144000/design-expression-and-evaluation-of-novel-chimeric-protein-constructed-from-colorectal-tumor-associated-antigen
#6
Ziba Veisi Malekshahi, Jafar Amani, Babak Negahdari, Masoumeh Rajabibazi, Walead Ebrahimizadeh
There were 134,000 new diagnosis and 49,000 deaths in 2016 due to colorectal cancer. Similar to most cancers, early diagnosis increases the chance of successful treatment. Detection of tumor-associated antigens or the immune response against such markers is one of the most common methods of diagnosis. In that regard, we aimed to design and express a chimeric protein from the most common tumor-associated antigens in colorectal cancer and assess its ability to detect the immune response in comparison with the parental tumor-associated antigens in patient's sera...
November 16, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/29143881/rmce-based-insect-cell-platform-to-produce-membrane-proteins-captured-on-hiv-1-gag-virus-like-particles
#7
João Vidigal, Bárbara Fernandes, Mafalda M Dias, Marco Patrone, António Roldão, Manuel J T Carrondo, Paula M Alves, Ana P Teixeira
Conformationally complex membrane proteins (MPs) are therapeutic targets in many diseases, but drug discovery has been slowed down by the lack of efficient production tools. Co-expression of MPs with matrix proteins from enveloped viruses is a promising approach to obtain correctly folded proteins at the surface of virus-like particles (VLPs), preserving their native lipidic environment. Here, we implemented a site-specific recombinase-mediated cassette exchange (RMCE) strategy to establish a reusable HIV-1 Gag-expressing insect cell line for fast production of target MPs on the surface of Gag-VLPs...
November 16, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29143761/antibody-cross-reactivity-between-porcine-cytomegalovirus-pcmv-and-human-herpesvirus-6-hhv-6
#8
Uwe Fiebig, Angela Holzer, Daniel Ivanusic, Elena Plotzki, Hartmut Hengel, Frank Neipel, Joachim Denner
Porcine cytomegalovirus (PCMV) infection is widely prevalent among pigs, and PCMV is one of the viruses which may be transmitted during xenotransplantation using pig cells, tissues, or organs. While human cytomegalovirus (HCMV) is a major risk factor for allotransplantation, it is still unclear whether PCMV is able to infect human cells or pose a risk for xenotransplantation. Previously, it was shown that transmission of PCMV after pig kidney to non-human primate transplantations resulted in a significantly reduced survival time of the transplanted organ...
October 28, 2017: Viruses
https://www.readbyqxmd.com/read/29143081/generation-in-yeast-and-antigenic-characterization-of-hepatitis-e-virus-capsid-protein-virus-like-particles
#9
Martynas Simanavicius, Paulius Lukas Tamosiunas, Rasa Petraityte-Burneikiene, Reimar Johne, Rainer G Ulrich, Aurelija Zvirbliene, Indre Kucinskaite-Kodze
Hepatitis E is a globally distributed human disease caused by hepatitis E virus (HEV). In Europe, it spreads through undercooked pork meat or other products and with blood components through transfusions. There are no approved or golden standard serologic systems for HEV diagnostics. Commercially available HEV tests often provide inconsistent results which may differ among the assays. In this study, we describe generation in yeast and characterization of HEV genotype 3 (HEV-3) and rat HEV capsid proteins self-assembled into virus-like particles (VLPs) and the development of HEV-specific monoclonal antibodies (MAbs)...
November 15, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29142300/structure-and-immunogenicity-of-pre-fusion-stabilized-human-metapneumovirus-f-glycoprotein
#10
Michael B Battles, Vicente Más, Eduardo Olmedillas, Olga Cano, Mónica Vázquez, Laura Rodríguez, José A Melero, Jason S McLellan
Human metapneumovirus (hMPV) is a frequent cause of bronchiolitis in young children. Its F glycoprotein mediates virus-cell membrane fusion and is the primary target of neutralizing antibodies. The inability to produce recombinant hMPV F glycoprotein in the metastable pre-fusion conformation has hindered structural and immunological studies. Here, we engineer a pre-fusion-stabilized hMPV F ectodomain and determine its crystal structure to 2.6 Å resolution. This structure reveals molecular determinants of strain-dependent acid-induced fusion, as well as insights into refolding from pre- to post-fusion conformations...
November 16, 2017: Nature Communications
https://www.readbyqxmd.com/read/29140753/a-mutated-recombinant-subunit-vaccine-protects-mice-and-guinea-pigs-against-botulinum-type-a-intoxication
#11
Chi Ho Yu, Dong Hyun Song, Jun Young Choi, Hae Eun Joe, Woo Hyeon Jeong, Gyeung Haeng Hur, Young Kee Shin, Seong Tae Jeong
Botulinum neurotoxins (BoNTs) are the most potent toxins to mammals. A toxoid vaccine was previously used for prevention of botulinum intoxication; however, this vaccine is no longer available. Currently, no approved botulinum vaccines are available from the Food and Drug Administration (FDA). Recently, a recombinant host cell receptor-binding subunit created for use as a potential vaccine completed phase 2 clinical trials. The current study designed a vaccine candidate against BoNT type A (BoNT/A) using a structural design...
November 15, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/29140390/a-carboxylated-graphene-nanodisks-glucose-oxidase-nanotags-and-mn-cds-tio2-matrix-based-dual-signal-amplification-strategy-for-ultrasensitive-photoelectrochemical-detection-of-tumor-markers
#12
Jie Song, Shuo Wu, Xinlan Yang, Jingli Yuan
Graphene nanodisks with good conductivity and plenty of edge sites were synthesized to load glucose oxidase (GRD-GOD) and coupled with a Mn(2+) doped CdS quantum dot (QD) modified TiO2 electrode (CdS:Mn/TiO2) for a highly sensitive photoelectrochemical (PEC) immunoassay. The specific immune-recognition behaviour can bring the GRD-GOD labelled antigen into the antibody immobilized CdS:Mn/TiO2 interface and dramatically enhance the photocurrent response via a dual signal amplification strategy. First, graphene nanodisks with a strong electron transfer capacity can improve the conductivity of both the insulating protein layers and the CdS:Mn/TiO2 matrix, thus facilitating the regeneration of trapped carriers and hot electrons in the CdS:Mn QD films and enhancing the PEC performance...
November 15, 2017: Analyst
https://www.readbyqxmd.com/read/29138852/anti%C3%A2-il%C3%A2-39-il%C3%A2-23p19-ebi3-polyclonal-antibodies-ameliorate-autoimmune-symptoms-in-lupus%C3%A2-like-mice
#13
Xiaoqian Wang, Yu Zhang, Zhiding Wang, Xiaoling Liu, Gaizhi Zhu, Gencheng Han, Guojiang Chen, Chunmei Hou, Tianxiao Wang, Beifen Shen, Yan Li, He Xiao, Ning Ma, Renxi Wang
The interleukin (IL)‑12 family cytokines have been examined as therapeutic targets in the treatment of several autoimmune diseases. Our previous study showed that a novel IL‑12 family cytokine, IL‑39 (IL‑23p19/Ebi3) mediates inflammation in lupus‑like mice. In the present study, the effect of anti‑mouse IL‑39 polyclonal antibodies on autoimmune symptoms in lupus‑like mice was investigated. Rabbit anti‑mouse IL‑39 polyclonal antibodies were produced by immunization with recombinant mouse IL‑39, and purified using protein A chromatography...
November 14, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29138320/structural-basis-for-antibody-recognition-of-the-nanp-repeats-in-plasmodium-falciparum-circumsporozoite-protein
#14
David Oyen, Jonathan L Torres, Ulrike Wille-Reece, Christian F Ockenhouse, Daniel Emerling, Jacob Glanville, Wayne Volkmuth, Yevel Flores-Garcia, Fidel Zavala, Andrew B Ward, C Richter King, Ian A Wilson
Acquired resistance against antimalarial drugs has further increased the need for an effective malaria vaccine. The current leading candidate, RTS,S, is a recombinant circumsporozoite protein (CSP)-based vaccine against Plasmodium falciparum that contains 19 NANP repeats followed by a thrombospondin repeat domain. Although RTS,S has undergone extensive clinical testing and has progressed through phase III clinical trials, continued efforts are underway to enhance its efficacy and duration of protection. Here, we determined that two monoclonal antibodies (mAbs 311 and 317), isolated from a recent controlled human malaria infection trial exploring a delayed fractional dose, inhibit parasite development in vivo by at least 97%...
November 14, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29138085/in-silico-design-cloning-expression-and-immunologic-evaluation-of-ed-fusion-protein-of-nt-h-influenzae
#15
Ava Behrouzi, Saeid Bouzari, Mana Oloomi, Abolfaz Fateh, Farzam Vaziri, Parviz Afrough, Amir Amanzadeh, Seyed Davar Siadat
Infections due to nontypeable Haemophilus influenzae (NTHi) are important causes of child mortality throughout the world. Given the lack of effective vaccines for these strains and the spread and prevalence of these infections in the world, it is necessary to design novel vaccine candidates against these strains. D and E proteins are conserved membrane-specific lipoproteins among encapsulated and non-encapsulated H. influenza strains, which, according to the exposure surface and conservation degree between both strains, can be considered as vaccine candidates suitable for studies...
November 11, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/29137976/proteomic-analysis-of-cho-cell-lines-producing-high-and-low-quantities-of-a-recombinant-antibody-before-and-after-selection-with-methotrexate
#16
Ruth Hausmann, Ivana Chudobová, Holger Spiegel, Stefan Schillberg
High levels of recombinant protein production in Chinese hamster ovary (CHO) cells can be achieved by amplification of transgenes using the dihydrofolate reductase/methotrexate (DHFR/MTX) system. With the aim to identify predictive markers enabling the preselection of suitable high producing clones we investigated the impact of MTX-based gene amplification on two CHO cells lines producing different levels of a human monoclonal antibody by carrying out a comparative proteome analysis. The difference in antibody yield between the high and low producer was 15-fold before and 245-fold after MTX selection...
November 11, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/29137362/an-optimized-hmgb1-expressed-by-recombinant-rabies-virus-enhances-immunogenicity-through-activation-of-dendritic-cells-in-mice
#17
Zhao Wang, Qian Liang, Yajing Zhang, Jie Yang, Mingming Li, Kunlun Wang, Min Cui, Huanchun Chen, Zhen F Fu, Ling Zhao, Ming Zhou
Rabies remains an important public health threat, killing approximately 59,000 people worldwide annually, most of which are from the developing countries of Africa and Asia where dog rabies are endemic. Therefore, developing an affordable and efficacious vaccine for dog-mediated rabies control is needful in these countries. Our previous studies indicated that over-expression of granulocyte-macrophage colony stimulating factor (GM-CSF) or macrophage inflammatory protein-1 (MIP-1α or CCL3) by recombinant rabies virus (rRABV) could enhance the immunogenicity by activating dendritic cells (DCs)...
October 13, 2017: Oncotarget
https://www.readbyqxmd.com/read/29134676/a-safety-and-immunogenicity-study-of-a-novel-subunit-plague-vaccine-in-cynomolgus-macaques
#18
Li Liu, Dong Wei, Zhe Qu, Li Sun, Yufa Miao, Yanwei Yang, Jinbiao Lu, Weixin Du, Bingxiang Wang, Bo Li
Plague has led to millions of deaths in history and outbreaks continue to the present day. The efficacy limitations and safety concerns of the existing killed whole cell and live-attenuated vaccines call for the development of new vaccines. In this study, we evaluated the immunogenicity and safety of a novel subunit plague vaccine, comprising native F1 antigen and recombinant V antigen. The cynomolgus macaques in low- and high-dose vaccine groups were vaccinated at weeks 0, 2, 4 and 6, at dose levels of 15 μg F1 + 15 μg rV and 30 μg F1 + 30 μg rV respectively...
November 14, 2017: Journal of Applied Toxicology: JAT
https://www.readbyqxmd.com/read/29134458/animal-cell-expression-systems
#19
M Butler, Ing U Reichl
The glycan profile of therapeutic recombinant proteins such as monoclonal antibodies is a critical quality attribute, which affects the efficacy of the final product. The cellular glycosylation process during protein expression is dependent upon a number of factors such as the availability of substrates in the media, the intracellular content of nucleotide sugars, and the enzyme repertoire of the host cells. In order to control the variability of glycosylation it is important to understand the critical process parameters and their acceptable range of values to enable reproducible production of proteins with a predetermined glycan profile providing the desired biological function or therapeutic effect...
October 3, 2017: Advances in Biochemical Engineering/biotechnology
https://www.readbyqxmd.com/read/29132995/comparative-functional-potency-of-dna-vaccines-encoding-plasmodium-falciparum-transmission-blocking-target-antigens-pfs48-45-and-pfs25-administered-alone-or-in-combination-by-in-vivo-electroporation-in-rhesus-macaques
#20
Dibyadyuti Datta, Geetha P Bansal, Brooke Grasperge, Dale S Martin, Mario Philipp, Dietlind Gerloff, Barry Ellefsen, Drew Hannaman, Nirbhay Kumar
Antibodies recognizing conformational epitopes in Pfs48/45, an antigen expressed on the surface of Plasmodium falciparum gametes and zygotes, have firmly established Pfs48/45 as a promising transmission blocking vaccine (TBV) candidate. However, it has been difficult to reproducibly express Pfs48/45 in a variety of recombinant expression systems. The goal of our studies was to evaluate functional immunogenicity of Pfs48/45 using DNA vaccine format in rhesus macaques. An additional goal was to ensure that when used in combination with another malarial antigen, specific immunity to both antigens was not compromised...
November 10, 2017: Vaccine
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