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vlp analysis

Bin Wang, Yujie Wang, Dylan A Frabutt, Xihe Zhang, Xiaoyu Yao, Dan Hu, Zhuo Zhang, Chaonan Liu, Shimin Zheng, Shi-Hua Xiang, Yong-Hui Zheng
The Ebola virus (EBOV) trimeric envelope glycoprotein (GP) precursors are cleaved into the receptor-binding GP1 and the fusion-mediating GP2 subunits and incorporated into virions to initiate infection. GP1 and GP2 form heterodimers, which have 15 or 2 N-glycosylation sites (NGSs), respectively. Here, we investigated the mechanism of how N-glycosylation contributes to GP expression, maturation, and function. As reported before, we found that although GP1 NGSs are not critical, the two GP2 NGSs, N563 and N618, are essential to the GP function...
February 14, 2017: Journal of Biological Chemistry
Anne-Marie C Andersson, Mafalda Resende, Ali Salanti, Morten A Nielsen, Peter J Holst
The malaria parasite Plasmodium falciparum presents antigens on the infected erythrocyte surface that bind human receptors expressed on the vascular endothelium. The VAR2CSA mediated binding to a distinct chondroitin sulphate A (CSA) is a crucial step in the pathophysiology of placental malaria and the CSA binding region of VAR2CSA has been identified as a promising vaccine target against placental malaria. Here we designed adenovirus encoded virus-like particles (VLP) by co-encoding Simian Immunodeficiency Virus (SIV) gag and VAR2CSA...
January 25, 2017: Vaccine
Petra Steppert, Daniel Burgstaller, Miriam Klausberger, Andres Tover, Eva Berger, Alois Jungbauer
The rapid quantification of enveloped virus-like particles (VLPs) requires orthogonal methods to obtain reliable results. Three methods-nanoparticle tracking analysis (NTA), size-exclusion HPLC (SE-HPLC) with UV detection, and detection with multi-angle light scattering (MALS)-for quantification of enveloped VLPs have been compared, and the lower and upper limits of detection and quantification have been evaluated. NTA directly counts the enveloped VLPs, and a particle number is obtained with a lower limit of detection (LLOD) of 1...
January 4, 2017: Journal of Chromatography. A
Kuniaki Nerome, Sayaka Matsuda, Kenichi Maegawa, Shigeo Sugita, Kazumichi Kuroda, Kazunori Kawasaki, Reiko Nerome
In this study, we aimed to quantitatively compare the increased production of three H7 influenza virus-like particle (VLP) haemagglutinin (HA) with the use of a codon-optimized single HA gene in silkworm pupae. Recombinant baculovirus (Korea H7-BmNPV) could produce 0.40 million HA units per pupa, corresponding to 1832μg protein. The yield of the HA produced in larva was estimated to be approximately 0.31 million HA units per larva, and there were no significant differences between the three HA proteins. We could establish efficient recovery system of HA production in larvae and pupae with the use of three cycles sonication methods...
February 1, 2017: Vaccine
Che-Ming Jack Hu, Chu-Yang Chien, Ming-Tsan Liu, Zih-Syun Fang, Sui-Yuan Chang, Rong-Huay Juang, Shih-Chung Chang, Hui-Wen Chen
BACKGROUND: Human infection with avian influenza A virus (H7N9) was first reported in China in March 2013. Since then, hundreds of cases have been confirmed showing severe symptoms with a high mortality rate. The virus was transmitted from avian species to humans and has spread to many neighboring areas, raising serious concerns over its pandemic potential. Towards containing the disease, the goal of this study is to prepare a virus-like particle (VLP) that consists of hemagglutinin (HA), neuraminidase (NA) and matrix protein 1 (M1) derived from the human isolate A/Taiwan/S02076/2013(H7N9) for potential vaccine development...
January 7, 2017: BMC Biotechnology
Mengying Cai, Cheng Wang, Yufeng Li, Hongjing Gu, Sujing Sun, Yueqiang Duan, Chengcai Lai, Keyu Wang, Xiaolan Yang, Li Xing, Peirui Zhang, Zhaohai Wang, Shaogeng Zhang, Xiaodong Guo, Shubing Liu, Yigang Tong, Xiliang Wang, Penghui Yang
Respiratory syncytial virus (RSV) is a leading cause of lower respiratory infection in infants and children, but there is still no licensed vaccine available. In this report, we developed virus-like particle (VLP) vaccines based on the Bac-to-Bac baculovirus expression system, consisting of an influenza virus matrix (M1) protein and the RSV fusion protein (F) or glycoprotein (G). These RSV VLPs were identified by western blot analysis and electron microscopy. Female BALB/c mice immunized intranasally (i.n.) with RSV-F VLPs, RSV-G VLPs, or both showed viral-specific antibody responses against RSV...
January 2017: Acta Biochimica et Biophysica Sinica
Nopporn Chutiwitoonchai, Lowela Siarot, Eri Takeda, Tatsuo Shioda, Motoki Ueda, Yoko Aida
HIV-1 budding requires interaction between Gag and cellular TSG101 to initiate viral particle assembly and release via the endosomal sorting complexes required for transport (ESCRT) pathway. However, some reports show that overexpression of TSG101 inhibits virus release by disruption of Gag targeting process. Since a HIV-1 accessory protein, Vpr binds to Gag p6 domain at the position close to the binding site for TSG101, whether Vpr implicates TSG101 overexpression effect has not been investigated. Here, we found that Vpr abrogates TSG101 overexpression effect to rescue viral production...
2016: PloS One
Asha Rani, Ravi Ranjan, Halvor S McGee, Ahmed Metwally, Zahraa Hajjiri, Daniel C Brennan, Patricia W Finn, David L Perkins
Recent studies have established that the human urine contains a complex microbiome, including a virome about which little is known. Following immunosuppression in kidney transplant patients, BK polyomavirus (BKV) has been shown to induce nephropathy (BKVN), decreasing graft survival. In this study we investigated the urine virome profile of BKV+ and BKV- kidney transplant recipients. Virus-like particles were stained to confirm the presence of VLP in the urine samples. Metagenomic DNA was purified, and the virome profile was analyzed using metagenomic shotgun sequencing...
September 16, 2016: Scientific Reports
Suneth S Perera, Bin Wang, Arturo Damian, Wayne Dyer, Li Zhou, Viviane Conceicao, Nitin K Saksena
HIV p24 is an extracellular HIV antigen involved in viral replication. Falling p24 antibody responses are associated with clinical disease progression and their preservation with non-progressive disease. Stimulation of p24 antibody production by immunization to delay progression was the basis of discontinued p24 vaccine. We studied a therapy-naive HIV+ man from Sydney, Australia, infected in 1988. He received the HIV-p24-virus like particle (VLP) vaccine in 1993, and continues to show vigorous p24 antigen responses (>4% p24-specific CD4⁺ T cells), coupled with undetectable plasma viremia...
2016: Microarrays
Aida Llauró, Benjamin Schwarz, Ranjit Koliyatt, Pedro J de Pablo, Trevor Douglas
Virus-like particles (VLPs) provide engineering platforms for the design and implementation of protein-based nanostructures. These capsids are comprised of protein subunits whose precise arrangement and mutual interactions determine their stability, responsiveness to destabilizing environments, and ability to undergo morphological transitions. The precise interplay between subunit contacts and the overall stability of the bulk capsid population remains poorly resolved. Approaching this relationship requires a combination of techniques capable of accessing nanoscale properties, such as the mechanics of individual capsids, and bulk biochemical procedures capable of interrogating the stability of the VLP ensemble...
September 27, 2016: ACS Nano
Moushira M Mostafa, Medhat A Al-Ghobashy, Faten A Fathalla, Maissa Y Salem
A sandwich-type ELISA was optimized and validated to determine the in-vitro relative potency of the four-component prophylactic Human papillomavirus (HPV) vaccine. The vaccine contains the non-infectious virus like particles (VLP) corresponding to HPV Types 6, 11, 16 and 18. A modification of the desorption step required to release the VLPs from the aluminum adjuvant was carried out. Samples were incubated with citrate buffer for two hours at 37 °C instead of overnight incubation at room temperature. Assay validation was then carried out according to ICH guidelines...
November 2016: Biologicals: Journal of the International Association of Biological Standardization
Manuela Cabiati, Benedetta Svezia, Marco Matteucci, Luca Botta, Angela Pucci, Mauro Rinaldi, Chiara Caselli, Vincenzo Lionetti, Silvia Del Ry
Osteopontin (OPN) is a phosphoglycoprotein of cardiac extracellular matrix and it is still poorly defined whether its expression changes in failing heart of different origin. The full-length OPN-a and its isoforms (OPN-b, OPN-c) transcriptomic profile were evaluated in myocardium of patients with dilated or ischemic cardiomyopathy (DCM n = 8; LVEF% = 17.5±3; ICM n = 8; LVEF% = 19.5±5.2) and in auricle of valvular patients (VLP n = 5; LVEF%≥50), by Real-time PCR analysis. OPN-a and thrombin mRNA levels resulted significantly higher in DCM compared to ICM patients (DCM:31...
2016: PloS One
Min Li, Xin Wang, Lu Cao, Zhijie Lin, Minxi Wei, Mujin Fang, Shaowei Li, Jun Zhang, Ningshao Xia, Qinjian Zhao
Human papillomavirus (HPV) 6 is a human pathogen which causes genital warts. Recombinant virus-like particle (VLP) based antigens are the active components in prophylactic vaccines to elicit functional antibodies. The binding and functional characteristics of a panel of 15 murine monoclonal antibodies (mAbs) against HPV6 was quantitatively assessed. Elite conformational indicators, recognizing the conformational epitopes, are also elite viral neutralizers as demonstrated with their viral neutralization efficiency (5 mAbs with neutralization titer below 4ng/mL) in a pseudovirion (PsV)-based system...
August 17, 2016: Vaccine
Jessica L Martin, Sheng Cao, Jose O Maldonado, Wei Zhang, Louis M Mansky
UNLABELLED: The Gag protein is the main retroviral structural protein, and its expression alone is usually sufficient for production of virus-like particles (VLPs). In this study, we sought to investigate-in parallel comparative analyses-Gag cellular distribution, VLP size, and basic morphological features using Gag expression constructs (Gag or Gag-YFP, where YFP is yellow fluorescent protein) created from all representative retroviral genera: Alpharetrovirus, Betaretrovirus, Deltaretrovirus, Epsilonretrovirus, Gammaretrovirus, Lentivirus, and Spumavirus We analyzed Gag cellular distribution by confocal microscopy, VLP budding by thin-section transmission electron microscopy (TEM), and general morphological features of the VLPs by cryogenic transmission electron microscopy (cryo-TEM)...
September 15, 2016: Journal of Virology
Huiming Huang, Lukuan Hou, Huayue Li, Yanhong Qiu, Jianhua Ju, Wenli Li
BACKGROUND: Actinomycete genome sequencing has disclosed a large number of cryptic secondary metabolite biosynthetic gene clusters. However, their unavailable or limited expression severely hampered the discovery of bioactive compounds. The whiB-like (wbl) regulatory genes play important roles in morphological differentiation as well as secondary metabolism; and hence the wblA so gene was probed and set as the target to activate cryptic gene clusters in deepsea-derived Streptomyces somaliensis SCSIO ZH66...
2016: Microbial Cell Factories
Elissa Meites, Pamina M Gorbach, Beau Gratzer, Gitika Panicker, Martin Steinau, Tom Collins, Adam Parrish, Cody Randel, Mark McGrath, Steven Carrasco, Janell Moore, Akbar Zaidi, Jim Braxton, Peter R Kerndt, Elizabeth R Unger, Richard A Crosby, Lauri E Markowitz
BACKGROUND: Gay, bisexual, and other men who have sex with men (MSM) are at high risk for human papillomavirus (HPV) infection; vaccination is recommended for US males, including MSM through age 26 years. We assessed evidence of HPV among vaccine-eligible MSM and transgender women to monitor vaccine impact. METHODS: During 2012-2014, MSM aged 18-26 years at select clinics completed a computer-assisted self-interview regarding sexual behavior, human immunodeficiency virus (HIV) status, and vaccinations...
September 1, 2016: Journal of Infectious Diseases
Petr Chlanda, Elena Mekhedov, Hang Waters, Cindi L Schwartz, Elizabeth R Fischer, Rolf J Ryham, Fredric S Cohen, Paul S Blank, Joshua Zimmerberg
Influenza A virus hemagglutinin (HA) changes conformation and drives membrane fusion of viral and endosomal membrane at low pH. Membrane fusion proceeds through an intermediate called hemifusion(1,2). For viral fusion the hemifusion structures are not determined(3). Here, influenza virus-like particles (VLP)(4) carrying wild-type (WT) HA or HA hemifusion mutant G1S(5) and liposome mixtures were studied at low pH by Volta phase plate (VPP) cryo-electron tomography (cET) which improves signal-to-noise ratio close to focus...
2016: Nature Microbiology
Young-Tae Lee, Eun-Ju Ko, Youri Lee, Yu-Na Lee, Zhen Bian, Yuan Liu, Sang-Moo Kang
UNLABELLED: An integrin-associated protein CD47, which is a ligand for the inhibitory receptor signal regulatory protein α, is expressed on B and T cells, as well as on most innate immune cells. However, the roles of CD47 in the immune responses to viral infection or vaccination remain unknown. We investigated the role of CD47 in inducing humoral immune responses after intranasal infection with virus or immunization with influenza virus-like particles (VLPs). Virus infection or vaccination with VLPs containing hemagglutinin from A/PR8/34 influenza virus induced higher levels of antigen-specific IgG2c isotype dominant antibodies in CD47-deficient (CD47KO) mice than in wild-type (WT) mice...
August 1, 2016: Journal of Virology
Makoto Ujike, Cheng Huang, Kazuya Shirato, Shinji Makino, Fumihiro Taguchi
The cytoplasmic tails of some coronavirus (CoV) spike (S) proteins contain an endoplasmic reticulum retrieval signal (ERRS) that can retrieve S proteins from the Golgi to the endoplasmic reticulum (ER); this process is thought to accumulate S proteins at the CoV budding site, the ER-Golgi intermediate compartment (ERGIC), and to facilitate S protein incorporation into virions. However, we showed previously that porcine epidemic diarrhoea CoV S proteins lacking the ERRS were efficiently incorporated into virions, similar to the original virus...
August 2016: Journal of General Virology
Gi-Ja Lee, Su-Hwa Lee, Young Ju Lee, Fu-Shi Quan
Sf9 cells (a clonal isolate of Spodoptera frugiperda Sf21 cells) are commonly used to generate recombinant virus-like particles (VLPs). For VLPs generation, Sf9 cells are infected with recombinant baculoviruses (rBV) expressing desired proteins. During rBV infections, Sf9 cells have changes in cell diameters and surface structures. In this study, for the first time, we investigated nanostructural changes of Sf9 cells using atomic force microscopy (AFM) during VLPs generation containing Toxoplasma gondii rhoptry protein 18 (ROP 18)...
November 2016: Scanning
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