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Recombinant antibody

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https://www.readbyqxmd.com/read/28350826/expression-purification-of-metallothionein-genes-from-freshwater-crab-sinopotamon-yangtsekiense-and-development-of-an-anti-metallothionein-elisa
#1
Jian Yang, Hui Sun, Hao Zhang, Hui Zhou
Using the phoA-fusion technology, the recombinant metallothionein (MT) from freshwater crab (Sinopotamon yangtsekiense) has been successfully produced in Escherichia coli. MT purified from the bacterial suspension showed one polypeptide with a molecular weight of 7 kDa by tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE). Western-blotting confirmed the polypeptides had a specific reactivity with mouse polyclonal MT anti-serum. Based on the purified MT and MT anti-serum, the reaction parameters for an enzyme-linked immunosorbent assay (ELISA) were developed...
2017: PloS One
https://www.readbyqxmd.com/read/28346811/improved-soluble-scfv-elisa-screening-approach-for-antibody-discovery-using-phage-display-technology
#2
Mohammad R Tohidkia, Maryam Sepehri, Shirin Khajeh, Jaleh Barar, Yadollah Omidi
Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the "biopanning" process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs...
March 1, 2017: SLAS Discov
https://www.readbyqxmd.com/read/28346319/effect-of-combination-antiretroviral-therapy-on-hiv-1-specific-antibody-dependent-cellular-cytotoxicity-responses-in-subtype-b-and-subtype-c-infected-cohorts
#3
Vijaya Madhavi, Archana Kulkarni, Ashwini Shete, Wen Shi Lee, Milla R Mclean, Anne B Kristensen, Manisha Ghate, Bruce D Wines, P Mark Hogarth, Matthew S Parsons, Anthony Kelleher, David A Cooper, Janaki Amin, Sean Emery, Madhuri Thakar, Stephen J Kent
BACKGROUND: There is growing interest in immune therapies to clear the latent HIV-1 after combination antiretroviral therapy (cART). There is limited information on the effect of cART on antibody-dependent cellular cytotoxicity (ADCC) and no studies have directly compared ADCC in HIV-1 subtype B- and subtype C-infected subjects. The effect of improving immunocompetence on ADCC to influenza also remains unexplored. METHODS: The effect of cART on HIV-1- and influenza-specific ADCC was analyzed in two cohorts (39 subtype B- and 47 subtype C-infected subjects) before and after two years of cART...
March 23, 2017: Journal of Acquired Immune Deficiency Syndromes: JAIDS
https://www.readbyqxmd.com/read/28346102/phage-display-derived-ligand-for-mucosal-transcytotic-receptor-gp-2-promotes-antigen-delivery-to-m-cells-and-induces-antigen-specific-immune-response
#4
Inam Ullah Khan, Jiansheng Huang, Rui Liu, Jingbo Wang, Jun Xie, Naishuo Zhu
Successful oral immunization depends on efficient delivery of antigens (Ags) to the mucosal immune induction site. Glycoprotein-2 (GP-2) is an integral membrane protein that is expressed specifically on M cells within follicle-associated epithelium (FAE) and serves as transcytotic receptor for luminal Ags. In this study, we selected peptide ligands against recombinant human GP-2 by screening a phage display library and evaluated their interaction with GP-2 in vitro and ex vivo. Selected peptides were conjugated to the C-terminal of enhanced green fluorescence protein (EGFP) and evaluated for their ability to induce an immune response in mice...
February 1, 2017: SLAS Discov
https://www.readbyqxmd.com/read/28346009/reactions-with-antisera-and-pathological-effects-of-staphylococcus-aureus-gamma-toxin-in-the-cornea
#5
Michael A Bierdeman, Angela M Torres, Armando R Caballero, Aihua Tang, Richard J O'Callaghan
PURPOSE: This study analyzed the toxicity of purified gamma-toxin from Staphylococcus aureus and the protectiveness of antisera to gamma-toxin in the rabbit cornea. MATERIALS AND METHODS: Gamma-toxin was purified from cultures of alpha-toxin deficient S. aureus strain Newman Δhla. Antisera to native gamma-toxin (Hlg) were produced in rabbits. These antisera and a commercial polyclonal antibody to recombinant HlgB (rHlgB) were analyzed for specificity and toxin neutralization...
March 27, 2017: Current Eye Research
https://www.readbyqxmd.com/read/28345468/surface-bound-galectin-4-regulates-gene-transcription-and-secretion-of-chemokines-in-human-colorectal-cancer-cell-lines
#6
U Subrahmanyeswara Rao, Prema S Rao
One long-term complication of chronic intestinal inflammation is the development of colorectal cancer. However, the mechanisms linking inflammation to the colorectal tumorigenesis are poorly defined. Previously, we have demonstrated that galectin-4 is predominantly expressed in the luminal epithelia of the gastrointestinal tract, and its loss of expression plays a key role in the colorectal tumorigenesis. However, the mechanism by which galectin-4 regulates inflammation-induced tumorigenesis is unclear. Here, we show that galectin-4 secreted by the colorectal cancer cell lines was bound to the cell surface...
March 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/28345293/detection-of-zaire-ebolavirus-in-swine-assay-development-and-optimization
#7
B S Pickering, B Collignon, G Smith, P Marszal, G Kobinger, H M Weingartl
Ebolaviruses (family Filoviridae, order Mononegavirales) cause often fatal, haemorrhagic fever in primates including humans. Pigs have been identified as a species susceptible to Reston ebolavirus (RESTV) infection, with indicated transmission to humans in the Philippines; however, their role during Ebola outbreaks in Africa needs to be clarified. To perform surveillance studies, detection of ebolavirus requires a prerequisite validation of viral RNA and antibody detection methods in swine samples. These diagnostic tests also need to be suitable for deployment to low-level containment laboratories...
March 27, 2017: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/28345289/severe-bleeding-diatheses-in-an-elderly-patient-with-combined-type-autoantibody-against-factor-xiii-a-subunit-novel-approach-to-the-diagnosis-and-classification-of-anti-factor-xiii-antibodies
#8
M Kun, N Szuber, É Katona, K Pénzes, A Bonnefoy, B Bécsi, F Erdődi, G E Rivard, L Muszbek
INTRODUCTION: Acquired factor XIII (FXIII) deficiency due to autoantibody is a rare, severe bleeding diathesis. Its laboratory diagnosis and classification represents a difficult task. AIM: Introduction of novel approaches into the diagnosis and characterization of anti-FXIII autoantibody and demonstration of their use in the diagnosis of a patient with autoimmune FXIII deficiency. METHODS: Factor XIII activity, FXIII antigen levels and the titre of anti-FXIII-A antibody were monitored throughout the course of the disease...
March 26, 2017: Haemophilia: the Official Journal of the World Federation of Hemophilia
https://www.readbyqxmd.com/read/28344998/low-dose-liver-targeted-gene-therapy-for-pompe-disease-enhances-therapeutic-efficacy-of-ert-via-immune-tolerance-induction
#9
Sang-Oh Han, Giuseppe Ronzitti, Benjamin Arnson, Christian Leborgne, Songtao Li, Federico Mingozzi, Dwight Koeberl
Pompe disease results from acid α-glucosidase (GAA) deficiency, and enzyme replacement therapy (ERT) with recombinant human (rh) GAA has clinical benefits, although its limitations include the short half-life of GAA and the formation of antibody responses. The present study compared the efficacy of ERT against gene transfer with an adeno-associated viral (AAV) vector containing a liver-specific promoter. GAA knockout (KO) mice were administered either a weekly injection of rhGAA (20 mg/kg) or a single injection of AAV2/8-LSPhGAA (8 × 10(11) vector genomes [vg]/kg)...
March 17, 2017: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/28344769/application-of-antibodies-to-recombinant-heat-shock-protein-70-in-immunohistochemical-diagnosis-of-mycobacterium-avium-subspecies-paratuberculosis-in-tissues-of-naturally-infected-cattle
#10
Julius Boniface Okuni, David Patrick Kateete, Moses Okee, Anna Nanteza, Moses Joloba, Lonzy Ojok
BACKGROUND: Detection of Mycobacterium avium subspecies paratuberculosis (MAP) infection is key to the control of Johne's disease. Immunohistochemistry is one of the methods of detection of MAP infection in tissues. However, unavailability of commercial antibodies that can detect the organism is a limiting factor for the use of immunohistochemistry. This study was aimed at developing an immunohistochemistry method to diagnose MAP in infected tissues using antibodies against MAP recombinant heat shock protein 70kd...
2017: Irish Veterinary Journal
https://www.readbyqxmd.com/read/28343774/immunogenicity-and-safety-of-a-high-dose-hepatitis-b-vaccine-among-patients-receiving-methadone-maintenance-treatment-a-randomized-double-blinded-parallel-controlled-trial
#11
Jing Shi, Yongliang Feng, Linying Gao, Dan Feng, Tian Yao, Shan Shi, Yawei Zhang, Xiaofeng Liang, Suping Wang
BACKGROUND AND AIMS: To explore whether the immunization with high-dose (60μg) hepatitis B vaccines in patients receiving methadone maintenance treatment (MMT) could yield a superior protection against hepatitis B infection than did the standard dose (20μg). METHODS: We conducted a randomized, double-blinded, parallel-controlled trial in MMT patients. Patients with serologically negative hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (anti-HBs) were randomized in a ratio of 1:1 to receive three intramuscular injections of 20μg or 60μg recombinant hepatitis B vaccine at months 0, 1, and 6...
March 23, 2017: Vaccine
https://www.readbyqxmd.com/read/28343748/b-cells-expressing-the-iga-receptor-fcrl4-participate-in-the-autoimmune-response-in-patients-with-rheumatoid-arthritis
#12
Khaled Amara, Elizabeth Clay, Lorraine Yeo, Daniel Ramsköld, Julia Spengler, Natalie Sippl, James A Cameron, Lena Israelsson, Philip J Titcombe, Caroline Grönwall, Ilfita Sahbudin, Andrew Filer, Karim Raza, Vivianne Malmström, Dagmar Scheel-Toellner
The clinical efficacy of B cell targeting therapies highlights the pathogenic potential of B cells in inflammatory diseases. Expression of Fc Receptor like 4 (FcRL4) identifies a memory B cell subset, which is enriched in the joints of patients with rheumatoid arthritis (RA) and in mucosa-associated lymphoid tissue. The high level of RANKL production by this B cell subset indicates a unique pathogenic role. In addition, recent work has identified a role for FcRL4 as an IgA receptor, suggesting a potential function in mucosal immunity...
March 24, 2017: Journal of Autoimmunity
https://www.readbyqxmd.com/read/28343398/an-anti-programmed-death-1-antibody-a%C3%AF-pd-1-fusion-protein-that-self-assembles-into-a-multivalent-and-functional-apd-1-nanoparticle
#13
Peng Zhao, Djordje Atanackovic, Shuyun Dong, Hideo Yagita, Xiao He, Mingnan Chen
Cancer immune checkpoint therapy has achieved remarkable clinical successes in various cancers. However, current immune checkpoint inhibitors block the checkpoint of not only the immune cells that are important to cancer therapy but also the immune cells that are irrelevant to the therapy. Such an indiscriminate blockade limits the efficacy and causes the autoimmune toxicity of the therapy. It might be beneficial to use a carrier to target immune checkpoint inhibitors to cancer-reactive immune cells. Here, we explore a method to load the inhibitors into carriers...
March 25, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28343172/novel-immunoassays-for-detection-of-cuzd1-autoantibodies-in-serum-of-patients-with-inflammatory-bowel-diseases
#14
Sofia Farkona, Antoninus Soosaipillai, Panagiota Filippou, Christos Liaskos, Dimitrios P Bogdanos, Eleftherios P Diamandis, Ivan M Blasutig
BACKGROUND: Pancreatic autoantibodies (PABs) are detected in patients with inflammatory bowel disease (IBD). Their prevalence is higher in Crohn's disease (CrD) than in ulcerative colitis (UC). Glycoprotein 2 (GP2) and, more recently, CUB and zona pellucida-like domain-containing protein 1 (CUZD1) have been identified as target autoantigens of PAB. The clinical utility of CUZD1 autoantibodies has only recently been assessed by indirect immunofluorescence (IIF) assays. In this study, we developed and validated novel immunoassays for the detection of CUZD1 autoantibodies...
March 27, 2017: Clinical Chemistry and Laboratory Medicine: CCLM
https://www.readbyqxmd.com/read/28343102/acoustic-and-hybrid-3d-printed-electrochemical-biosensors-for-the-real-time-immunodetection-of-liver-cancer-cells-hepg2
#15
Samar Damiati, Seta Küpcü, Martin Peacock, Christoph Eilenberger, Mazin Zamzami, Ishtiaq Qadri, Hani Choudhry, Uwe B Sleytr, Bernhard Schuster
This study presents an efficient acoustic and hybrid three-dimensional (3D)-printed electrochemical biosensors for the detection of liver cancer cells. The biosensors function by recognizing the highly expressed tumor marker CD133, which is located on the surface of liver cancer cells. Detection was achieved by recrystallizing a recombinant S-layer fusion protein (rSbpA/ZZ) on the surface of the sensors. The fused ZZ-domain enables immobilization of the anti-CD133 antibody in a defined manner. These highly accessible anti-CD133 antibodies were employed as a sensing layer, thereby enabling the efficient detection of liver cancer cells (HepG2)...
March 21, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28342851/site-specific-photocoupling-of-pbpa-mutated-scfv-antibodies-for-use-in-affinity-proteomics
#16
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/28342665/recombinant-newcastle-disease-virus-expressing-the-infectious-bronchitis-virus-s1-gene-protects-chickens-against-newcastle-disease-virus-and-infectious-bronchitis-virus-challenge
#17
Ran Zhao, Junfeng Sun, Tianming Qi, Wen Zhao, Zongxi Han, Xiaopu Yang, Shengwang Liu
The recombinant LaSota strain expressing a chimeric IBV S1 gene (rLaSota-S1) was constructed with the S1 gene of the LX4 type IBV ck/CH/LDL/091022. The expression of the S1 protein was detected by an indirect immunofluorescence assay and Western blotting. The rLaSota-S1 strain was slightly attenuated, and its growth dynamics were similar to that of the parental LaSota strain. Vaccination of specific pathogen-free chickens with the rLaSota-S1 strain induced NDV hemagglutination inhibition antibodies, and it protected chickens from challenge with virulent NDV...
March 22, 2017: Vaccine
https://www.readbyqxmd.com/read/28342316/comparison-of-cytomegalovirus-cmv-specific-neutralization-capacity-of-hyperimmunoglobulin-hig-versus-standard-intravenous-immunoglobulin-ivig-preparations-impact-of-cmv-igg-normalization
#18
Matthias Stefan Schampera, Katrin Schweinzer, Harald Abele, Karl Oliver Kagan, Reinhild Klein, Ingo Rettig, Gerhard Jahn, Klaus Hamprecht
BACKGROUND: Based on a non-randomized study of Nigro et al. (2005) the intravenous administration of hyperimmunoglobulins (HIGs) is applied frequently to women with primary CMV-infection as "off-label use" in Germany. OBJECTIVES: In order to describe their CMV-specific neutralization-capacity in vitro, we analyzed the HIG preparations Cytotect(®), and Cytogam(®) as well as the standard intravenous immunoglobulins (IVIG) Octagam(®), Gamunex(®), Kiovig(®). STUDY DESIGN: We performed short-term cell-free CMV neutralization assays (CFNT) and long-term cell-adapted neutralization-plaque-reduction assays (PRANT)...
March 16, 2017: Journal of Clinical Virology: the Official Publication of the Pan American Society for Clinical Virology
https://www.readbyqxmd.com/read/28342171/expressing-anti-hiv-vrc01-antibody-using-the-murine-igg1-secretion-signal-in-pichia-pastoris
#19
Rochelle Aw, Paul F McKay, Robin J Shattock, Karen M Polizzi
The use of the recombinant expression platform Pichia pastoris to produce pharmaceutically important proteins has been investigated over the past 30 years. Compared to mammalian cultures, expression in P. pastoris is cheaper and faster, potentially leading to decreased costs and process development times. Product yields depend on a number of factors including the secretion signal chosen for expression, which can influence the host cell response to recombinant protein production. VRC01, a broadly neutralising anti-HIV antibody, was expressed in P...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28341898/functional-block-of-il-17-cytokine-promotes-bone-healing-by-augmenting-foxo1-and-atf4-activity-in-cortical-bone-defect-model
#20
M Dixit, K B Singh, R Prakash, D Singh
We determine the effect of interleukin (IL)-17 neutralizing antibody on new bone regeneration. Anti-IL-17 antibody promoted new bone regeneration in cortical bone defect model by augmenting FOXO1 and ATF4 activity thereby decreasing oxidative stress. Our study demonstrates the bone healing and regeneration potential of neutralizing IL-17antibody in osteoporotic fractures. INTRODUCTION: The immune system plays important role in the fracture healing process. However, fracture healing is prolonged in disorders associated with systemic inflammation...
March 24, 2017: Osteoporosis International
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