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spike protein

Steffen Großhans, Susanna Suhm, Jürgen Hubbuch
Preparative protein precipitation is known as a cost-efficient and easy-to-use alternative to chromatographic purification steps. This said, at the moment, there is no process for monoclonal antibodies (mAb) on the market, although especially polyethylene glycol-induced precipitation has shown great potential. One reason might be the highly complex behavior of each component of a crude feedstock during the precipitation process. For different investigated mAbs, significant variations in the host cell protein (HCP) reduction are observed...
March 18, 2019: Bioprocess and Biosystems Engineering
Toru Matsu-Ura, Hideki Shirakawa, Kenichi G N Suzuki, Akitoshi Miyamoto, Kotomi Sugiura, Takayuki Michikawa, Akihiro Kusumi, Katsuhiko Mikoshiba
In most species, fertilization induces Ca2+ transients in the egg. In mammals, the Ca2+ rises are triggered by phospholipase Cζ (PLCζ) released from the sperm; IP3 generated by PLCζ induces Ca2+ release from the intracellular Ca2+ store through IP3 receptor, termed IP3 -induced Ca2+ release. Here, we developed new fluorescent IP3 sensors (IRIS-2s) with the wider dynamic range and higher sensitivity (Kd = 0.047-1.7 μM) than that we developed previously. IRIS-2s employed green fluorescent protein and Halo-protein conjugated with the tetramethylrhodamine ligand as fluorescence resonance energy transfer (FRET) donor and acceptor, respectively...
March 18, 2019: Scientific Reports
David Bartolomé-Martín, Ignacio Ibáñez, Dolores Piniella, Elena Martínez-Blanco, Sara G Pelaz, Francisco Zafra
Dopamine and glutamate transporters (DAT and GLT-1, respectively) share some biophysical characteristics, as both are secondary active carriers coupled to electrochemical ion gradients. In order to identify common or specific components of their respective proteomes, we performed a proximity labelling assay (BioID) in the hippocampal cell line HT22. While most of the identified proteins were specific for each transporter (and will be analyzed elsewhere), we detected two membrane proteins in the shared interactome of GLT-1 and DAT: the transmembrane protein 263 (Tmem263) and the potassium channel protein Kv7...
March 15, 2019: Neuropharmacology
Rebecca Ye, Sirisha Kundrapu, Stanton L Gerson, James J Driscoll, Rose Beck, Naveed Ali, Ola Landgren, Willem VanHeeckeren, George Luo, Nicolaus Kroger, Paolo Caimi, Marcos De Lima, Ehsan Malek
BACKGROUND: High-dose chemotherapy and autologous stem cell transplantation (ASCT) are integral components of the overall treatment for patients with multiple myeloma (MM) aged ≤ 65 years. The emergence of oligoclonal immunoglobulin bands (ie, immunoglobulins differing from those originally identified at diagnosis [termed clonal isotype switch (CIS)]) has been reported in patients with MM after high-dose chemotherapy followed by autologous stem cell transplantation. However, the clinical relevance and the correlation with immune reconstitution remains unclear...
January 3, 2019: Clinical Lymphoma, Myeloma & Leukemia
Chen Guoning, Guo Pengqi, Wang Yan, Wang Lu, Shu Hua, Li Yunzhe, Jing Wanghui, Chang Chun, Fu Qiang
Immunoassay as a primary analytical tool is widely applied in the field of analysis. However, biological antibodies used in the routine immunoassay exhibit high cost and poor stability. Herein, in this study, a biomimetic ELISA method using molecularly imprinted polymers as a alternative to antibodies was developed. The molecularly imprinted polymers were prepared through dopamine polymerization using Fe3 O4 nanoparticles as magnetic nuclei, bovine serum albumin as a template, dopamine as a functional monomer and crosslinking agent...
June 1, 2019: Talanta
Jason W Westerbeck, Carolyn E Machamer
Coronaviruses (CoVs) assemble by budding into the lumen of the early Golgi prior to exocytosis. The small CoV envelope (E) protein plays roles in assembly, virion release, and pathogenesis. CoV E has a single hydrophobic domain (HD), is targeted to Golgi membranes, and has cation channel activity in vitro The E protein from the avian infectious bronchitis virus (IBV) has dramatic effects on the secretory system, which require residues in the HD. Mutation of the HD of IBV E in a recombinant virus background results in impaired growth kinetics, impaired release of infectious virions, accumulation of IBV spike (S) protein on the plasma membrane when compared IBV WT infected cells, and aberrant cleavage of IBV S on virions...
March 13, 2019: Journal of Virology
Kelly Niermans, Jan Woyzichovski, Nina Kröncke, Rainer Benning, Ronald Maul
Edible insects as additional food and/or feed source may represent one important component to solve the problem of food security for a growing human population. Especially for covering the rising demand for protein of animal origin, seven insect species currently allowed as feed constituents in the European Union are gaining more interest. However, before considering insects such as yellow mealworm larvae (Tenebrio molitor) as suitable for, e.g. human consumption, the possible presence and accumulation of contaminants must be elucidated...
March 13, 2019: Mycotoxin Research
Nicola Brady, Emily L O'Reilly, Christopher McComb, Alastair I Macrae, P David Eckersall
In cattle, the serum protein haptoglobin (Hp) is a major acute phase protein (APP) that rises in concentration over a thousand fold following stimulation by pro-inflammatory cytokines. As such, this APP is a valuable biomarker for infection, inflammation and trauma in cattle. The assay for bovine Hp is becoming more commonplace in clinical pathology and in experimental studies when a biomarker of innate immunity is required. The most widely used assay for Hp utilises its binding to haemoglobin (Hp-Hb binding assay), which at low pH enables the preservation of the native peroxidase activity in the haemoglobin...
2019: Comparative Clinical Pathology
Mengru Guo, Shushuai Yi, Yanbing Guo, Shuang Zhang, Jiangting Niu, Kai Wang, Guixue Hu
Porcine epidemic diarrhea caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious disease in newborn piglets. The spike (S) protein is the surface glycoprotein of PEDV, which can induce specific neutralization antibodies and is a candidate antigen for vaccination attempts. In our study, the S1 region of PEDV strain CH/JLDH/2016 spike gene was inserted into the Lactococcus lactis expression vector, pNZ8149, resulting in recombinant plasmid pNZ8149-S1, and the immunogenicity of recombinant L. lactis pNZ8149-S1/NZ3900 was evaluated in mice...
March 11, 2019: Viral Immunology
Mengyao Wang, Honglin Yang, Yue Wu, Zhifeng Fu
A dual sites affinity protocol was developed for fluorescent analysis of Staphylococcus aureus (S. aureus) by employing daptomycin and immunoglobulin G (IgG) as the recognition elements. Pig IgG immobilized on microplate was employed as the first recognition element to capture S. aureus owing to the fact that the Fc segment of mammal IgG can selectively bind with protein A on the surface of the target bacteria. Meanwhile, fluorescein isothiocyanate-conjugated daptomycin was employed as the second recognition element as well as the signal tracer for the target bacteria utilizing the binding capability of daptomycin to Gram-positive bacteria...
February 19, 2019: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
Pearlson Prashanth Austin Suthanthiraraj, Ashis Kumar Sen
Early diagnosis of dengue biomarkers by employing a technology that is less labor- and time-intensive and offers higher sensitivity and lower limits of detection would find great significance in the developing world. Here, we report the development of a biosensor that exploits the localized surface plasmon resonance (LSPR) effect of silver nanostructures, created via thermal annealing of thin metal film, to detect dengue NS1 antigen, which appears as early as the onset of infection. The biosensor integrates membrane-based blood-plasma separation to develop lab-on-chip device that facilitates rapid diagnosis (within 30 min) of dengue NS1 antigen from a small volume (10 µL) of whole blood...
February 21, 2019: Biosensors & Bioelectronics
Laila Ali Layqah, Shimaa Eissa
The Middle East respiratory syndrome corona virus (MERS-CoV) is highly pathogenic. An immunosensor for the determination of MERS-CoV is described here. It is based on a competitive assay carried out on an array of carbon electrodes (DEP) modified with gold nanoparticles. Recombinant spike protein S1 was used as a biomarker for MERS CoV. The electrode array enables multiplexed detection of different CoVs. The biosensor is based on indirect competition between free virus in the sample and immobilized MERS-CoV protein for a fixed concentration of antibody added to the sample...
March 7, 2019: Mikrochimica Acta
Leo Brueggeman, Morgan L Sturgeon, Russell M Martin, Andrew J Grossbach, Yasunori Nagahama, Angela Zhang, Mathew A Howard, Hiroto Kawasaki, Shu Wu, Robert A Cornell, Jacob J Michaelson, Alexander G Bassuk
Objective: Epilepsy treatment falls short in ~30% of cases. A better understanding of epilepsy pathophysiology can guide rational drug development in this difficult to treat condition. We tested a low-cost, drug-repositioning strategy to identify candidate epilepsy drugs that are already FDA-approved and might be immediately tested in epilepsy patients who require new therapies. Methods: Biopsies of spiking and nonspiking hippocampal brain tissue from six patients with unilateral mesial temporal lobe epilepsy were analyzed by RNA-Seq...
February 2019: Annals of Clinical and Translational Neurology
Mark A J Bayliss, Katarina Rigdova, Mathew Kyriakides, Sally Grier, Andew M Lovering, Hannah Williams, David C Griffith, Alasdair MacGowan
New treatments are urgently required to treat infections caused by multi-drug resistant Acinetobacter baumanni,. To address this need, a new formulation of Minocin®, (minocycline for injection) has been developed that allows for higher doses of minocycline to be administered. Phase 1 clinical trials were conducted in healthy volunteers to assess the safety and pharmacokinetics (PK) of this new formulation at higher doses. In order to generate PK data, novel, selective and simple HPLC-MS/MS based assays were developed and validated for the determination of minocycline (MC) in human plasma and urine...
February 26, 2019: Journal of Pharmaceutical and Biomedical Analysis
Naeem Khan, Asghari Bano, M D Ali Babar
The present study was aimed to investigate the effects of plant growth promoting rhizobacteria (PGPR) and plant growth regulators (PGRs) on the physiology and yield of wheat grown in less fertile sandy soil. The isolated PGPR strains were identified by 16S-rRNA gene sequencing as Planomicrobium chinense (P1), Bacillus cereus (P2) and Pseudomonas fluorescens (P3). Wheat varieties (Galaxy-13 and Pak-2013) differing in sensitivity to drought were soaked in fresh cultures of bacterial isolates and the PGRs (salicylic acid and putrescine) were sprayed at 150 mg/L on seedlings at three leaf stage...
March 6, 2019: Archives of Microbiology
Kim M Bouwman, Mattias Delpont, Frederik Broszeit, Renaud Berger, Erik A W S Weerts, Marie-Noëlle Lucas, Maxence Delverdier, Sakhia Belkasmi, Andreas Papanikolaou, Geert-Jan Boons, Jean-Luc Guérin, Robert P de Vries, Mariette F Ducatez, Monique H Verheije
Guinea fowl coronavirus (GfCoV) causes fulminating enteritis that can result in a daily death rate of 20% in guinea fowl flocks. Here we studied GfCoV diversity and evaluated its phenotypic consequences. Over the period 2014-2016, affected guinea fowl flocks were sampled in France and avian coronavirus presence was confirmed by PCR on intestinal content and immunohistochemistry of intestinal tissue. Sequencing revealed 89% amino acid identity between the viral attachment protein S1 of GfCoV/2014 and the previously identified GfCoV/2011...
March 6, 2019: Journal of Virology
Li-Sha Wang, Meng-Di Zhang, Xue Tao, Yun-Feng Zhou, Xin-Min Liu, Rui-Le Pan, Yong-Hong Liao, Qi Chang
l-Tryptophan (Trp) metabolites and related neurotransmitters play crucial roles in physiological functions, and their imbalances are implicated in the pathology of depression, Alzheimer's disease and other diseases. Measurement of Trp metabolites and related neurotransmitters possesses a great potential to elucidate the disease mechanisms and evaluate the outcomes of therapeutic interventions. A simple, rapid, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of Trp, l-kynurenine (Kyn), kynurenic acid (Kyna), 3-hydroxykynurenine (3-HK), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE), l-glutamic acid (Glu), γ-aminobutyric acid (GABA) and acetylcholine (ACh) in mice serum and the brain tissues in a single chromatographic run...
February 22, 2019: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Manmeet Singh, Abhinoy Kishore, Dibyajyoti Maity, Punnepalli Sunanda, Bankala Krishnarjuna, Sreeparna Vappala, Srinivasarao Raghothama, Lawrence C Kenyon, Debnath Pal, Jayasri Das Sarma
Fusion peptides (FP) in spike proteins are key players mediating early events in cell-to-cell fusion, vital for intercellular viral spread. A proline residue located at the central FP region has often been suggested to have a distinctive role in this fusion event. The spike glycoprotein from strain RSA59 (PP) of mouse hepatitis virus (MHV) contains two central, consecutive prolines in the FP. Here, we report that deletion of one of these proline residues, resulting in RSA59 (P), significantly affected neural cell syncytia formation and viral titers post-infection in vitro...
March 1, 2019: Journal of Biological Chemistry
Nai-Chi Chen, Masato Yoshimura, Naoyuki Miyazaki, Hong-Hsiang Guan, Phimonphan Chuankhayan, Chien-Chih Lin, Shao-Kang Chen, Pei-Ju Lin, Yen-Chieh Huang, Kenji Iwasaki, Atsushi Nakagawa, Sunney I Chan, Chun-Jung Chen
Shrimp nodaviruses, including Penaeus vannamei (PvNV) and Macrobrachium rosenbergii nodaviruses (MrNV), cause white-tail disease in shrimps, with high mortality. The viral capsid structure determines viral assembly and host specificity during infections. Here, we show cryo-EM structures of T  = 3 and T  = 1 PvNV-like particles (PvNV-LPs), crystal structures of the protrusion-domains (P-domains) of PvNV and MrNV, and the crystal structure of the ∆N-ARM-PvNV shell-domain (S-domain) in T  = 1 subviral particles...
2019: Communications Biology
Min Sun Kim, Ki Hong Kim
Vaccines based on viral replicon particles would be advantageous to induce immune responses compared to inactivated viruses in that they can infect host cells (only once) and can produce viral proteins in the infected cells like live viruses. Furthermore, as viral replicon particles are replication-defective, they are safer than live attenuated viruses. Previously, we had rescued viral hemorrhagic septicemia virus (VHSV) replicon particles lacking full ORF of G gene (rVHSV-ΔG). In the present study, to enhance the immunogenicity of VHSV replicon particles, we newly generated another form of VHSV replicon particles that can produce the transmembrane and C-terminal cytoplasmic region-deleted G protein in host cells (rVHSV-GΔTM), and compared the protective efficacy of rVHSV-GΔTM with that of rVHSV-ΔG through immunization of olive flounder (Paralichthys olivaceus)...
February 25, 2019: Fish & Shellfish Immunology
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