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https://www.readbyqxmd.com/read/28516782/exploring-site-specific-n-glycosylation-of-hek293-and-plant-produced-human-iga-isotypes
#1
Kathrin Göritzer, Daniel Maresch, Friedrich Altmann, Christian Obinger, Richard Strasser
The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. Here, monomeric IgA1, IgA2m(1) and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. All three IgA isotypes were purified and subjected to biophysical and biochemical characterization...
May 18, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28509333/altered-glycan-accessibility-on-native-immunoglobulin-g-complexes-in-early-rheumatoid-arthritis-and-its-changes-during-therapy
#2
Judith Stümer, Mona H C Biermann, Jasmin Knopf, Iryna Magorivska, Alf Kastbom, Anna Svärd, Christina Janko, Rostyslav Bilyy, Georg Schett, Christopher Sjöwall, Martin Herrmann, Luis E Muñoz
The goal of this study was to investigate the glycosylation profile of native IgG present in serum immune complexes in patients with rheumatoid arthritis (RA). To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins were employed. Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically-linked galactose/N-acetylgalactosamine (GalNac-L) were used...
May 16, 2017: Clinical and Experimental Immunology
https://www.readbyqxmd.com/read/28486931/accuracy-of-m2bpgi-compared-with-fibro-scan%C3%A2-in-analysis-of-liver-fibrosis-in-patients-with-hepatitis-c
#3
Hongqin Xu, Wenli Kong, Lei Liu, Xiumei Chi, Xiaomei Wang, Ruihong Wu, Xiuzhu Gao, Huan Wang, Limei Qu, Yue Qi, Yu Pan, Junqi Niu
BACKGROUND: Mac-2 Binding Protein Glycosylation isomer (M2BPGi) is a novel serological glyco-biomarker for staging liver fibrosis. Here, we aimed to evaluate the efficiency of serum M2BPGi in identifying liver fibrosis stages in Chinese patients with chronic hepatitis C infection. METHODS: Serum M2BPGi levels were evaluated in 680 patients with chronic hepatitis C and 164 healthy controls who underwent the Fibro Scan® test of liver fibrosis. The diagnostic accuracy of serum M2BPGi values was compared to that of other fibrosis markers, including Fibro Scan®, the aspartate transaminase to platelet ratio index (APRI), the fibrosis index based on four factors (FIB4), and the gamma-glutamyltranspeptidase to platelet ratio (GPR)...
May 10, 2017: BMC Gastroenterology
https://www.readbyqxmd.com/read/28466278/improving-antibody-based-cancer-therapeutics-through-glycan-engineering
#4
REVIEW
Xiaojie Yu, Michael J E Marshall, Mark S Cragg, Max Crispin
Antibody-based therapeutics has emerged as a major tool in cancer treatment. Guided by the superb specificity of the antibody variable domain, it allows the precise targeting of tumour markers. Recently, eliciting cellular effector functions, mediated by the Fc domain, has gained traction as a means by which to generate more potent antibody therapeutics. Extensive mutagenesis studies of the Fc protein backbone has enabled the generation of Fc variants that more optimally engage the Fcγ receptors known to mediate cellular effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and cellular phagocytosis...
May 2, 2017: BioDrugs: Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy
https://www.readbyqxmd.com/read/28460523/block-sequence-specific-glycopolypeptides-with-selective-lectin-binding-properties
#5
Cristina Lavilla, Gokhan Yilmaz, Veselina Uzunova, Richard Napier, C Remzi Becer, Andreas Heise
Glycopolypeptides with defined block sequences were prepared by sequential addition of two different N-carboxyanhydrides (NCAs), followed by selective deprotection and functionalization of predefined positions within the polypeptide backbone. The sequential arrangement of the galactose units and the block-sequence length have been systematically varied. All the glycopolypeptides have been obtained with a similar overall composition and comparable molecular weights. Circular dichroism measurements revealed some dependence of the secondary structure on the primary composition of the glycopolypeptides at physiological pH...
May 11, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28453257/one-filter-one-sample-and-the-n-and-o-glyco-proteo-me-toward-a-system-to-study-disorders-of-protein-glycosylation
#6
Kirsty Skeene, Matthew Walker, Graham Clarke, Ed Bergström, Paul Genever, Daniel Ungar, Jane Thomas-Oates
A method has been developed for release/isolation of O-glycans from glycoproteins in whole cell lysates for mass spectrometric analysis. Cells are lysed in SDS, which is then exchanged for urea and ammonium bicarbonate in a centrifugal filter, before treating with NH4OH to release O-glycans. Following centrifugation, O-glycans are recovered in the filtrate. Sonication achieves O-glycan release in 1 h. Combining the established protocol for filter-aided N-glycan separation, here optimized for enhanced PNGase F efficiency, with the developed O-glycan release method allows analysis of both N- and O-glycans from one sample, in the same filter unit, from 0...
May 10, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28439939/identification-of-a-polygalacturonase-cup-s-2-as-the-major-ccd-bearing-allergen-in-cupressus-sempervirens-pollen
#7
Youcef Shahali, Jean-Pierre Sutra, Christiane Hilger, Kyria Swiontek, Iman Haddad, Joelle Vinh, Laurence Guilloux, Denis Charpin, Hélène Sénéchal, Pascal Poncet
Since IgE glyco-epitopes, also referred to as cross-reactive carbohydrate determinants (CCDs), can share significant structural homologies between different plants, they are prone to extensive cross-reactivity among allergen pollen extracts. Here, cypress pollen allergens, especially a polygalacturonase (PG), were further characterized using double one dimensional electrophoresis (D1-DE). The presence of specific IgE directed against CCDs was investigated by bromelain IgE inhibition and concanavalin A binding assays using sera of cypress pollen sensitized patients...
April 25, 2017: Allergy
https://www.readbyqxmd.com/read/28427157/the-anti-tumor-efficacy-of-3c23k-a-glyco-engineered-humanized-anti-misrii-antibody-in-an-ovarian-cancer-model-is-mainly-mediated-by-engagement-of-immune-effector-cells
#8
Pauline Estupina, Alexandre Fontayne, Jean-Marc Barret, Nathalie Kersual, Olivier Dubreuil, Marion Le Blay, Alexandre Pichard, Marta Jarlier, Martine Pugnière, Maëva Chauvin, Thierry Chardès, Jean-Pierre Pouget, Emmanuel Deshayes, Alexis Rossignol, Toufik Abache, Christophe de Romeuf, Aurélie Terrier, Lucie Verhaeghe, Christine Gaucher, Jean-François Prost, André Pèlegrin, Isabelle Navarro-Teulon
Ovarian cancer is the leading cause of death in women with gynecological cancers and despite recent advances, new and more efficient therapies are crucially needed. Müllerian Inhibiting Substance type II Receptor (MISRII, also named AMHRII) is expressed in most ovarian cancer subtypes and is a novel potential target for ovarian cancer immunotherapy. We previously developed and tested 12G4, the first murine monoclonal antibody (MAb) against human MISRII. Here, we report the humanization, affinity maturation and glyco-engineering steps of 12G4 to generate the Fc-optimized 3C23K MAb, and the evaluation of its in vivo anti-tumor activity...
February 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28407380/n-glycan-maturation-mutants-in-lotus-japonicus-for-basic-and-applied-glycoprotein-research
#9
Carina T Pedersen, Ian Loke, Andrea Lorentzen, Sara Wolf, Manoj Kamble, Sebastian K Kristensen, David Munch, Simona Radutoiu, Edzard Spillner, Peter Roepstorff, Morten Thaysen-Andersen, Jens Stougaard, Svend Dam
Studies of protein N-glycosylation are important for answering fundamental questions on the diverse functions of glycoproteins in plant growth and development. Here we generated and characterised a comprehensive collection of Lotus japonicus LORE1 insertion mutants, each lacking the activity of one of the twelve enzymes required for normal N-glycan maturation in the glycosylation machinery. The inactivation of the individual genes resulted in altered N-glycan patterns as documented using mass spectrometry and glycan-recognising antibodies, indicating successful identification of null-mutations in the target glyco-genes...
April 13, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28400175/recombinant-plant-derived-human-ige-glycoproteomics
#10
Laura Montero-Morales, Daniel Maresch, Alexandra Castilho, Aysegül Turupcu, Kristina M Ilieva, Silvia Crescioli, Sophia N Karagiannis, Christian Lupinek, Chris Oostenbrink, Friedrich Altmann, Herta Steinkellner
The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2...
April 8, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28393916/production-and-glyco-engineering-of-immunomodulatory-helminth-glycoproteins-in-plants
#11
Ruud H P Wilbers, Lotte B Westerhof, Kim van Noort, Katja Obieglo, Nicole N Driessen, Bart Everts, Sonja I Gringhuis, Gabriele Schramm, Aska Goverse, Geert Smant, Jaap Bakker, Hermelijn H Smits, Maria Yazdanbakhsh, Arjen Schots, Cornelis H Hokke
Helminth parasites control host-immune responses by secreting immunomodulatory glycoproteins. Clinical trials and mouse model studies have demonstrated the potential of helminth-derived glycoproteins for the treatment of immune-related diseases, like allergies and autoimmune diseases. Studies are however hampered by the limited availability of native parasite-derived proteins. Moreover, recombinant protein production systems have thus far been unable to reconstitute helminth-like glycosylation essential for the functionality of some helminth glycoproteins...
April 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28388443/synthetic-glyco-o-sulfatome-for-profiling-of-human-natural-antibodies
#12
Galina Pazynina, Marina Sablina, Tatiana Ovchinnikova, Tatiana Tyrtysh, Svetlana Tsygankova, Alexander Tuzikov, Kira Dobrochaeva, Nadezhda Shilova, Nailia Khasbiullina, Nicolai Bovin
Our understanding of biological role of glycans O-sulfation remains at the level of beginners due to microheterogeneity, lability and other difficulties of exact structural assignment. Partially, problem of functional investigations, especially determination of glycoepitope specificity of carbohydrate-binding proteins could be solved with the help of synthetic glycans of certain structure. Here we summing up our synthetic efforts in creation of synthetic O-sulfatome, and bring together all the synthesized in our group sulfated glycans, both existing in nature, yet undiscovered but biochemically licit, and completely unnatural...
March 22, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/28362077/lactosylated-glycogen-nanoparticles-for-targeting-prostate-cancer-cells
#13
Quinn Alexander Besford, Marcin Wojnilowicz, Tomoya Suma, Nadja Bertleff-Zieschang, Frank Caruso, Francesca Cavalieri
Glyconanoparticles that exhibit multivalent binding to lectins are highly desirable for molecular recognition and therapeutic applications, for example. Herein, we explore the use of glycogen nanoparticles as a biosourced glyco-scaffold for engineering multivalent glyconanoparticles. Glycogen nanoparticles, a naturally occurring highly-branched polymer of glucose, was functionalized with lactose, achieved through copper(I)-catalyzed alkyne-azide cycloaddition chemistry, for targeted interaction with lectins ex situ and on prostate cancer cells...
March 31, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28344072/osteopontin-metallothionein-i-ii-interactions-in-experimental-autoimmunune-encephalomyelitis
#14
Hrvoje Jakovac, Tanja Grubić Kezele, Sandra Šućurović, Biserka Mulac-Jeričević, Biserka Radošević-Stašić
Osteopontin (OPN), an extracellular matrix (ECM) glyco-phosphoprotein, plays an important role in autoimmune-mediated demyelinating diseases, including multiple sclerosis and experimental autoimmune encephalomyelitis (EAE). As an integrin and CD44 binding protein it participates in bidirectional communication between the ECM and target cells and affects transduction pathways that maintain neuronal and immune cell homeostasis. Its biological activity is also heavily influenced by microenvironment, which stimulates the cleavage of OPN and changes its functions...
March 24, 2017: Neuroscience
https://www.readbyqxmd.com/read/28338836/a-novel-pathogenesis-of-inflammatory-bowel-disease-from-the-perspective-of-glyco-immunology
#15
Shinichiro Shinzaki, Hideki Iijima, Hironobu Fujii, Yoshihiro Kamada, Tetsuji Naka, Tetsuo Takehara, Eiji Miyoshi
Oligosaccharide modifications play an essential role in various inflammatory diseases and cancers, but their pathophysiologic roles, especially in inflammation, are not clear. Inflammatory bowel disease (IBD) is an intractable chronic inflammatory disorder with an unknown aetiology, and the number of patients with IBD is increasing throughout the world. Certain types of immunosuppressant drugs, such as corticosteroids, are effective for IBD, suggesting that immune function is closely associated with the pathophysiology of IBD...
May 1, 2017: Journal of Biochemistry
https://www.readbyqxmd.com/read/28337966/acid-fast-positive-and-acid-fast-negative-mycobacterium-tuberculosis-the-koch-paradox
#16
Catherine Vilchèze, Laurent Kremer
Acid-fast (AF) staining, also known as Ziehl-Neelsen stain microscopic detection, developed over a century ago, is even today the most widely used diagnostic method for tuberculosis. Herein we present a short historical review of the evolution of AF staining methods and discuss Koch's paradox, in which non-AF tubercle bacilli can be detected in tuberculosis patients or in experimentally infected animals. The conversion of Mycobacterium tuberculosis from an actively growing, AF-positive form to a nonreplicating, AF-negative form during the course of infection is now well documented...
March 2017: Microbiology Spectrum
https://www.readbyqxmd.com/read/28335280/pregnancy-vaccination-with-gold-glyco-nanoparticles-carrying-listeria-monocytogenes-peptides-protects-against-listeriosis-and-brain-and-cutaneous-associated-morbidities
#17
Ricardo Calderón-Gonzalez, Héctor Terán-Navarro, Elisabet Frande-Cabanes, Eva Ferrández-Fernández, Javier Freire, Soledad Penadés, Marco Marradi, Isabel García, Javier Gomez-Román, Sonsoles Yañez-Díaz, Carmen Álvarez-Domínguez
Listeriosis is a fatal infection for fetuses and newborns with two clinical main morbidities in the neonatal period, meningitis and diffused cutaneous lesions. In this study, we vaccinated pregnant females with two gold glyconanoparticles (GNP) loaded with two peptides, listeriolysin peptide 91-99 (LLO91-99) or glyceraldehyde-3-phosphate dehydrogenase 1-22 peptide (GAPDH1-22). Neonates born to vaccinated mothers were free of bacteria and healthy, while non-vaccinated mice presented clear brain affections and cutaneous diminishment of melanocytes...
August 19, 2016: Nanomaterials
https://www.readbyqxmd.com/read/28323046/centaurium-erythraea-methanol-extract-protects-red-blood-cells-from-oxidative-damage-in-streptozotocin-induced-diabetic-rats
#18
Miloš Đorđević, Mirjana Mihailović, Jelena Arambašić Jovanović, Nevena Grdović, Aleksandra Uskoković, Anja Tolić, Marija Sinadinović, Jovana Rajić, Danijela Mišić, Branislav Šiler, Goran Poznanović, Melita Vidaković, Svetlana Dinić
ETHNOPHARMACOLOGICAL RELEVANCE: Centaurium erythraea Rafn (CE) is a traditional medicinal herb in Serbia with antidiabetic, digestive, antipyretic and antiflatulent effects. AIM OF THE STUDY: To investigate the potential protective effects of the methanol extract of the aerial parts of CE against glyco-oxidative stress in red blood cells (RBCs) in rats with experimentally induced diabetes. MATERIAL AND METHODS: Diabetes was induced in Wistar rats by intraperitoneal (i...
March 16, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28318221/crystal-structure-of-a-homogeneous-igg-fc-glycoform-with-the-n-glycan-designed-to-maximize-the-antibody-dependent-cellular-cytotoxicity
#19
Chia-Lin Chen, Jen-Chi Hsu, Chin-Wei Lin, Chia-Hung Wang, Ming-Hung Tsai, Chung-Yi Wu, Chi-Huey Wong, Che Ma
N-glycosylation on IgG modulates Fc conformation and effector functions. An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure...
March 31, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28302960/development-of-chemical-and-chemo-enzymatic-glycosylations
#20
REVIEW
Shin-Ichiro Shoda
Glycosidic compounds are indispensable molecules in living systems. Biological phenomena such as cell wall formation, energy storage, and cell recognition strongly depend on the multi-functional characters of these substances. Development of highly regio- and stereoselective glycosylation reactions is necessary to provide sufficient amounts of specific compounds in basic research as well as for applications in industry. This review presents an overview of chemical and chemo-enzymatic glycosylations that have been developed during my forty-year academic career in the field of glyco-science...
2017: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences
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