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Recombinant baculovirus

Mary Lopez-Perez, Mads Delbo Larsen, Rafael Bayarri-Olmos, Paulina Ampomah, Liz Stevenson, Myriam Arévalo-Herrera, Sócrates Herrera, Lars Hviid
Clinical immunity to malaria is associated with acquisition of IgG specific for members of the Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family of clonally variant antigens on the surface of infected erythrocytes (IEs). The VAR2CSA subtype of PfEMP1 mediates IE binding in the placenta. VAR2CSA-specific IgG is normally acquired only after exposure to placental parasites. However, it was recently reported that men and children from Colombia often have high levels of functional VAR2CSA-specific IgG...
May 21, 2018: Infection and Immunity
Darien Kheder Ali Mohamed, Junzheng Du, Shandian Gao, Zhancheng Tian, Guorui Zhang, Dexuan Huang, Rongsheng Du, Biao Kang, Guangyuan Liu, Jianxun Luo, Hong Yin
Bluetongue virus (BTV), a vector-borne pathogen, is the causative agent of bluetongue disease in ruminants. In view of the recent emergence of BTV in regions previously known to be free from the disease and/or specific serotypes or strains, optimization of the currently available vaccination strategies to control the spread of vector-borne bluetongue is crucial. The main objective of the current study was to develop a subunit vaccine candidate targeting BTV-16, a strain previously isolated in China from sheep with obvious clinical signs...
June 2018: Veterinary Microbiology
Shuangyu Xie, Yiming Shi, Ruyue Gong, Wen Cui, Yanping Jiang, Min Liu, Li Wang, Han Zhou, Xinyuan Qiao, Yijing Li, Yigang Xu, Lijie Tang
Serious outbreaks of bluetongue, an arbovirus of domestic and wild ruminants caused by bluetongue virus serotypes (BTV), have occurred around the world. More than 27 distinct serotypes are recognized throughout the world. A new virus, BTV-25 (Toggenburg orbivirus [TOV]), was first detected in Switzerland, and has not yet been found in China. VP2 is an important outer shell protein that defines BTV serotypes and is, therefore, an ideal target antigen for serotype identification. To produce a monoclonal antibody against VP2 of BTV-25, the segment 2 gene was divided into three segments, cloned into pET-28a (+) and pMAL-c5X vectors, and the protein was expressed in E...
June 2018: Veterinary Microbiology
Wei Zhang, Wenlong Dai, Chao Zhang, Yu Zhou, Pei Xiong, Shuxia Wang, Xiaohua Ye, Qingwei Liu, Dongming Zhou, Zhong Huang
Hand, foot, and mouth disease (HFMD) is an infectious disease that mainly affects infants and children, causing considerable morbidity and mortality worldwide. HFMD is commonly caused by enterovirus 71 (EV71) and coxsackieviruses A16 (CVA16), A6 (CVA6), and A10 (CVA10). Formalin-inactivated EV71 vaccines are currently available in China; however, these vaccines fail to confer cross-protection against infections by other HFMD-causing enteroviruses, highlighting the necessity of developing a multivalent HFMD vaccine...
May 18, 2018: Emerging Microbes & Infections
Jiwoon Jeong, Changhoon Park, Seeun Kim, Su-Jin Park, Ikjae Kang, Kee Hwan Park, Chanhee Chae
A novel porcine circovirus type 2 (PCV2) peptide vaccine comprised of a consensus capsid (Cap) protein domain encoded by open reading frame 2 was developed to control PCV2 infection. The efficacy of the vaccine was evaluated against a commercial baculovirus-expressed recombinant PCV2 subunit vaccine based on the Cap protein. The amino acid sequence of this Cap protein was designed based on the alignment of amino acid sequences from different isolates from Europe, North America, and Asia. The vaccine was evaluated in either phosphate-buffered saline or adjuvanted with aluminum hydroxide, cobalt oxide, or liposome...
April 2018: Canadian Journal of Veterinary Research, Revue Canadienne de Recherche Vétérinaire
Xiaoyue Zhang, Keyan Xu, Yanmei Ou, Xiaodong Xu, Hongying Chen
BACKGROUND: The Baculovirus expression vector system (BEVS) is a transient expression platform for recombinant protein production in insect cells. Baculovirus infection of insect cells will shutoff host translation and induce apoptosis and lead to the termination of protein expression. Previous reports have demonstrated the enhancement of protein yield in BEVS using stable insect cell lines expressing interference RNA to suppress the expression of caspase-1. RESULTS: In this study, short-hairpin RNA (shRNA) expression cassettes targeting Spodoptera frugiperda caspase-1 (Sf-caspase-1) were constructed and inserted into an Autographa californica multiple nucleopolyhedrovirus (AcMNPV) vector...
May 2, 2018: BMC Biotechnology
Irene Melegari, Fulvio Marsilio, Federica Di Profio, Vittorio Sarchese, Ivano Massirio, Andrea Palombieri, Anna Rita D'Angelo, Gianvito Lanave, Georgia Diakoudi, Alessandra Cavalli, Vito Martella, Barbara Di Martino
Caliciviruses of the Sapovirus genus have been recently detected in dogs. Canine sapoviruses (SaVs) have been identified in the stools of young or juvenile animals with gastro-enteric disease at low prevalence (2.0-2.2%), but whether they may have a role as enteric pathogens and to which extent dogs are exposed to SaVs remains unclear. Here, we report the expression in a baculovirus system of virus like-particles (VLPs) of a canine SaV strain, the prototype virus Bari/4076/2007/ITA. The recombinant antigen was used to develop an enzyme-linked immunosorbent assay (ELISA)...
April 23, 2018: Virus Research
Bastian Franke, Christian Opitz, Shin Isogai, Anne Grahl, Leonildo Delgado, Alvar D Gossert, Stephan Grzesiek
Baculovirus-infected insect cells have become a powerful tool to express recombinant proteins for structural and functional studies by NMR spectroscopy. This article provides an introduction into the insect cell/baculovirus expression system and its use for the production of recombinant isotope-labeled proteins. We discuss recent advances in inexpensive isotope-labeling methods using labeled algal or yeast extracts as the amino acid source and give examples of advanced NMR applications for proteins, which have become accessible by this eukaryotic expression host...
April 23, 2018: Journal of Biomolecular NMR
Magalie Penaud-Budloo, Achille François, Nathalie Clément, Eduard Ayuso
Recombinant adeno-associated viral (rAAV) vectors have been used in more than 150 clinical trials with a good safety profile and significant clinical benefit in many genetic diseases. In addition, due to their ability to infect non-dividing and dividing cells and to serve as efficient substrate for homologous recombination, rAAVs are being used as a tool for gene-editing approaches. However, manufacturing of these vectors at high quantities and fulfilling current good manufacturing practices (GMP) is still a challenge, and several technological platforms are competing for this niche...
March 16, 2018: Molecular Therapy. Methods & Clinical Development
Md Nasimuzzaman, Johannes C M van der Loo, Punam Malik
Baculovirus has traditionally been used for the production of recombinant protein and vaccine. However, more recently, baculovirus is emerging as a promising vector for gene therapy application. Here, baculovirus is produced by transient transfection of the baculovirus plasmid DNA (bacmid) in an adherent culture of Sf9 cells. Baculovirus is subsequently expanded in Sf9 cells in a serum-free suspension culture until the desired volume is obtained. It is then purified from the culture supernatant using heparin affinity chromatography...
April 9, 2018: Journal of Visualized Experiments: JoVE
Yunwang Shen, Min Feng, Xiaofeng Wu
ORF40 (bm40) of the Bombyx mori nucleopolyhedrovirus (BmNPV) encodes a homologue of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) AC51 and is a highly conserved gene in sequenced alphabaculoviruses. To investigate the role of bm40 in the baculovirus infection cycle, a bm40 knockout BmNPV bacmid was constructed via homologous recombination in Escherichia coli. Western blotting analysis revealed that bm40 is a late gene during virus infection. Compared with wild-type and repair viruses, the knockout virus exhibited a single-cell infection phenotype...
April 19, 2018: Journal of General Virology
Ying Lou, Guorong Ji, Qin Liu, Pengbo Wang, Ruilin Zhang, Yuanxing Zhang, Xiaohong Liu
The human thyroid peroxidase (hTPO) is an essential enzyme for thyroid hormone biosynthesis and is expressed in thyroid cells. It is an autoantigen against which antibodies are found in the sera of patients with a number of autoimmune thyroid disorders. Overexpression of hTPO has been achieved using the baculovirus expression vector system (BEVS). However, it is produced largely in an aggregated form in the cell lysate fraction, which increases the complexity of protein extraction. In this study, to achieve improved secretory expression of hTPO via BEVS, a truncated recombinant hTPO protein (hTPOt) was engineered by replacing its original signal peptide (SP) in the N-terminal with five heterologous SPs...
April 12, 2018: Protein Expression and Purification
Ngonda Saasa, Masahiro Kajihara, George Dautu, Akina Mori-Kajihara, Shuetsu Fukushi, Yona Sinkala, Shigeru Morikawa, Aaron Mweene, Ayato Takada, Kumiko Yoshimatsu, Jiro Arikawa
The open reading frame of the nucleocapsid protein (NP) of Rift Valley fever virus (RVFV) strain MP12 was cloned and expressed in Vero E6 cells. The recombinant NP (rNP)-expressing cells were used as antigens for an indirect immunofluorescent antibody assay (IFA). The rNP-based IFA and RVFV-infected Vero E6 cell (authentic antigen)-based IFA showed similar IFA profiles with immune rabbit serum, which was prepared by immunization with rNP expressed using a baculovirus vector. A total of 942 traditional cattle sera obtained in five districts in Central, Southern, and Western provinces of Zambia were screened for anti-RVFV antibodies by the authentic antigen-based and rNP-based IFAs...
March 13, 2018: Vector Borne and Zoonotic Diseases
Kohei Tahara, Manami Kobayashi, Satoko Yoshida, Risako Onodera, Naoki Inoue, Hirofumi Takeuchi
In this study, we demonstrated that cationic liposomes with incorporated stearylamine (SA) inhibit viral infectivity without preloaded active pharmaceutical ingredients. Specifically, we correlated physiochemical properties of liposomes, such as zeta potentials and particle sizes, with virus infectivity using the BacMam™ reagent, which is based on recombinant baculovirus (BV). Compared with neutral or negatively-charged liposomes, SA liposomes suppressed BV infectivity in several mammalian cell lines, including A549 cells...
April 3, 2018: International Journal of Pharmaceutics
Z Yang, Y Hu, P Yuan, Y Yang, K Wang, L Y Xie, S L Huang, J Liu, L Ran, Z H Song
In the swine hepatitis E virus (HEV), open reading frame 2 (ORF2) is rich in antigenic determinants and neutralizing epitopes that could induce immune protection. We chose the Bac-to-Bac® Baculovirus Expression System to express fragments containing the critical neutralizing antigenic sites within the HEV ORF2 protein of pigs to obtain a recombinant baculovirus. The fragment of swine HEV ORF2 region (1198-1881bp) was cloned into vector pFastBacTM. A recombinant baculovirus, rBacmid-ORF2, was obtained after transposition and transfection...
March 2018: Polish Journal of Veterinary Sciences
Hee Geun Park, Kwang Sik Lee, Bo Yeon Kim, Hyung Joo Yoon, Yong Soo Choi, Kyung Yong Lee, Hu Wan, Jianhong Li, Byung Rae Jin
Honeybee (Apis mellifera) egg-yolk protein vitellogenin (Vg) plays roles in immunity, antioxidation, and life span beyond reproduction, but it also acts as an allergen Api m 12 in venom. Here we established antimicrobial and antioxidant roles of honeybee Vg in the body and venom. Using the cDNA encoding Vg identified from Asiatic honeybee (A. cerana) workers, recombinant A. cerana Vg (AcVg) protein of approximately 180 kDa was produced in baculovirus-infected insect cells. In A. cerana worker bees, AcVg was expressed in the fat body and venom gland and was present in the secreted venom...
April 2, 2018: Developmental and Comparative Immunology
Tsuyoshi Imasaki, Sabine Wenzel, Kentaro Yamada, Megan L Bryant, Yuichiro Takagi
The baculovirus expression vector system (BEVS) is becoming the method of choice for expression of many eukaryotic proteins and protein complexes for biochemical, structural and pharmaceutical studies. Significant technological advancement has made generation of recombinant baculoviruses easy, efficient and user-friendly. However, there is a tremendous variability in the amount of proteins made using the BEVS, including different batches of virus made to express the same proteins. Yet, what influences the overall production of proteins or protein complexes remains largely unclear...
2018: PloS One
Xi Wang, Ke Dong, Min Long, Fang Lin, Zhaowei Gao, Lin Wang, Zhe Zhang, Xi Chen, Ying Dai, Huiping Wang, Huizhong Zhang
Enterovirus 71 (EV71) is the most frequently detected causative agent in hand, foot, and mouth disease (HFMD) and is a serious threat to public health in the Asia-Pacific region. Many EV71 vaccines are under development worldwide, and although both inactivated virus vaccines and virus-like particles (VLPs) are considered to be effective, the main focus has been on inactivated EV71vaccines. There have been very few studies on EV71 VLPs. In this study, using a strategy based on HIV gag VLPs, we constructed a gag-VP1 fusion gene to generate a recombinant baculovirus expressing the EV71 structural protein VP1 together with gag, which was then used to infect TN5 cells to form VLPs...
March 27, 2018: Archives of Virology
Wang Xi, Dong Ke, Long Min, Wang Lin, Zuo Jiahui, Lin Fang, Gao Zhaowei, Zhang Zhe, Chen Xi, Zhang Huizhong
The cell surface glycoprotein Trop‑2 is overexpressed in various types of cancer, including in lung cancer, and has recently been used as an effective immunotherapeutic target. CD40 ligand (CD40L), a tumor necrosis factor superfamily member, is a promising immune adjuvant. Human immunodeficiency virus (HIV) gag‑based virus‑like particles (VLPs) are highly immunogenic, and foreign antigens can be incorporated onto their membrane envelope for cancer vaccine development. In the present study, a HIV gag‑based VLP strategy and Bac‑to‑Bac system were utilized to construct Trop‑2, CD40L and gag recombinant baculoviruses, which were then used to infect TN5 cells in order to form Trop‑2 VLPs or Trop‑2‑CD40L VLPs...
March 15, 2018: International Journal of Molecular Medicine
C W E Embregts, D Rigaudeau, L Tacchi, G P Pijlman, L Kampers, T Veselý, D Pokorová, P Boudinot, G F Wiegertjes, M Forlenza
We recently reported on a successful vaccine for carp against SVCV based on the intramuscular injection of a DNA plasmid encoding the SVCV glycoprotein (SVCV-G). This shows that the intramuscular (i.m.) route of vaccination is suitable to trigger protective responses against SVCV, and that the SVCV G-protein is a suitable vaccine antigen. Yet, despite the general success of DNA vaccines, especially against fish rhabdoviruses, their practical implementation still faces legislative as well as consumer's acceptance concerns...
March 19, 2018: Fish & Shellfish Immunology
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