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2 edition of Characterization and applications of recombinant HPV 16 E7 protein found in the catalog.

Characterization and applications of recombinant HPV 16 E7 protein

Adrianus Johannes Cornelis Maria Braspenning

Characterization and applications of recombinant HPV 16 E7 protein

by Adrianus Johannes Cornelis Maria Braspenning

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  • 11 Currently reading

Published by University of Leiden in [Leiden .
Written in English


Edition Notes

StatementAdrianus Johannes Cornelis Maria Braspenning.
Classifications
LC ClassificationsMLCM 2002/02258
The Physical Object
Pagination1 v. (unpaged) :
ID Numbers
Open LibraryOL478521M
LC Control Number98204687

  Daemen, Toos, et al. “Eradication of established HPV transformed tumours after immunisation with recombinant Semliki Forest virus expressing a fusion protein of E6 and E7.” Vaccine, (), vol. 21, pp. The changes in E7 mRNA levels are quickly reflected in E7 protein expression, since the half-life of the HPV E7 protein produced by recombinant baculoviruses in Sf insect cells at +37°C is less than 30

Cervical cancer is the second-most-common cause of malignancies in women worldwide, and the oncogenic activity of the human papilloma virus types (HPV) E7 protein has a crucial role in anogenital tumors. In this study, we have designed a therapeutic vaccine based on chitosan nanodelivery systems to deliver HPV E7 DNA vaccine, considered as a tumor specific antigen for immunotherapy of HPV Administration of an L. lactis strain displaying a cell wall-anchored HPV E7 antigen was significantly enhanced after coadministration of an L. lactis strain secreting IL protein, corroborating the hypothesis that the recombinant strain described here is a promising candidate for mucosal codelivery of proteins of medical interest. This

  The most detailed study on LAB-based vaccines concerns vaccination against human papillomavirus (HPV). HPV is the causative agent of cervical cancer and oncoprotein E7 produced in tumor cells is known to be an efficient antigen. There exist numerous studies on the development of vaccines against HPV that are based on using E7 [95–97]. The expression of vaccine antigens in lactic acid bacteria (LAB) is a safe and cost-effective alternative to traditional expression systems. In this study, we investigated i) the expression of Human papillomavirus type 16 (HPV) L1 major capsid protein in the model LAB Lactococcus lactis and ii) the ability of the resulting recombinant strain to produce either capsomer-or virus


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Characterization and applications of recombinant HPV 16 E7 protein by Adrianus Johannes Cornelis Maria Braspenning Download PDF EPUB FB2

Here, we describe a protocol for the successful isolation of RNA aptamers directed at the recombinant HPV E7 protein through the application of the SELEX method.

Once the nucleic acid sequence of a functional aptamer is determined, large amounts of the oligonucleotide can be produced and modified at low cost and high :// developed a HPV‐16 therapeutic vaccine candidate consisting of the HPV‐16 E7 oncoprotein fused to a peptide derived from the Limulus polyphemus antilipopolysaccharide factor (LALF 32‐51).

This is a small and hydrophobic peptide that can penetrate cell membranes and that has immunomodulatory properties (Granadillo et al.,). Aptamers were selected from a randomized oligonucleotide library using a modified SELEX method and recombinant HPV E7 protein.

Isolated aptamers were cloned and sequenced for in silico ://   HPV type 16 (HPV) is the most common HPV type associated with cervical cancer with an overall incidence of 50% in cervical tumors worldwide. The combined activities of the viral genes E6 and E7 cause genomic instability and are sufficient and necessary for cell immortalization and transformation 3, 4, :// Recombinant E7 from HPV purified to near homogeneity showed two species in gel filtration chromatography, one of these corresponding to a dimer with a molecular weight of 22 kDa, determined by /_Chemical_synthesis_of_the_HPV16_E7_protein.

E7 protein is a major oncogenic factor of human papillomaviruses (HPVs) that plays a key role in virus-associated human cervical carcinogenesis. To determine the biochemical properties of the E7 protein of high-risk HPV t the gene encoding the protein was cloned into a bacterial vector, pETa (+), to allow expression of HPVE7 as a thioredoxin (Trx) fusion protein in Because E7 (an early viral protein of HPV16) is continuously expressed by the target epithelial cell upon viral integration and cellular transformation, is highly conserved in amino acid sequence among HPV genotypes and is antigenic in man [11–13], immunotherapeutic strategies to enhance the endogenous response to this tumour-specific HPV 16 E6/E7 DNA was found only in OE‐E6/E7 but not in OE.

Human oviduct‐specific glycoprotein, estrogen receptors, and cytokeratin were found in both cell types. Both OE and OE‐E6/E7 possessed telomerase activities but the former had much lower ://   Cervical cancer is the sixth most common cancer in women worldwide and the leading cause of women’s death in developing countries.

Nearly all cervical cancers are associated with infection of the human papillomavirus (HPV). This sexually transmitted pathogen disrupts the cell cycle via two oncoproteins: E6 and E7.

Cells respond to E7-mediated   Development of a High Throughput Screen and Inhibitor Identification. A high throughput screen (HTS) for small molecule inhibitors of the E6–E6AP interaction was designed around a modified sandwich enzyme-linked immunosorbance assay (ELISA) (Figure (Figure1).

1).Full length HPV16 E6 was fused C terminally to glutathione-S-transferase (GST).To Protein & Peptide Letters publishes letters, original research papers, mini-reviews and guest edited issues in all important aspects of protein and peptide research, including structural studies, advances in recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, and drug design.

Manuscripts must have a significant element of novelty, timeliness ?journalID=ppl. Mice were also inoculated with amounts of purified recombinant (rec) HPV-E7 protein equivalent to those associated with the exosome inocula. Two weeks after the last inoculation, mice were challenged with 10 5 TC-1 cells, and tumor growth was monitored over time.

Mice inoculated with recombinant E7 have been sacrificed at day 27 due to their An inducible system to improve and stabilize the production of an extremely labile protein (E7 antigen of human papillomavirus type 16) was developed in the food‐grade bacterium Lactococcus lactis.

A protein carrier, the staphylococcal nuclease Nuc, was fused either to N/ or C‐termini of E7 protein, and the resulting hybrid proteins were In Human Papillomaviruses- (HPV-) associated carcinogenesis, continuous expression of the E6 oncoprotein supports its value as a potential target for the development of diagnostics and therapeutics for HPV cancer.

We previously reported that the I7 single-chain antibody fragment (scFv) specific for HPV16 E6, expressed as an intrabody by retroviral system, could inhibit Persistent infection by high risk HPV types (e.g. HPV, and ) is the main risk factor for development of cervical intraepithelial neoplasia and cervical cancer.

Tumor necrosis factor (TNF) is a key mediator of epithelial cell inflammatory response and exerts a potent cytostatic effect on normal or HPV16, but not on HPV18 immortalized ://   Then it was shown that the HPV 16 L1 capsid protein alone could self-assemble into VLPs. In these experiments, the yield of VLPs were low but the HPV 16 L1 gene used to generate the recombinant vectors was that of the prototype HPV 16 According to the HPV E7 protein sequence the calculated IEP of this protein ishowever, when this characteristic was measured with recombinant E7 protein produced in bacteria the IEP obtained was [48 Pahel G, Aulabaugh A, Short SA, et al.

Structural and functional characterization of the HPV16 E7 protein expressed in bacteria J Western blot analysis showing antibodies against HPV E7 protein after intranasal immunization with recombinant L. lactis NZE7-CWA. Cell extracts of E. coli expressing GST-E7 protein, used as antigen (+), were resolved in 12% SDS–PAGE gels, transferred onto polyvinylidene difluoride membrane and reacted with pooled sera from immunized ://   Identification of enolase in S.

thermophilus CGMCC To characterize whether the enolase from S. thermophilus could bind to microbial surface, an enolase encoding gene enoM was identified in the genome of S. thermophilus CGMCC It was bp in size and composed of amino acid residues.

Multiple-sequence alignment results showed that the deduced EnoM protein S. Peng, C. Trimble, L. Wu, D. Pardoll, C.-F. Hung, and T.-C.

Wu () HLA-DQB1*Restricted HPV E7 Peptide-specific CD4+ T Cell Immune Responses Correlate with Regression of HPVAssociated High Grade Squamous Intraepithelial Lesions Clinical Cancer Research.

SABBL has been tested as an immunomodulatory component of an HPV E7 recombinant protein subunit-based vaccine on an HPV-induced mouse cancer model. The SABBL E7-based subunit vaccine exhibited significant therapeutic efficacy by eradicating established tumors in approximately 70% of ://  To generate the recombinant construct (L1/E6/E7) encoding CTL-epitopes restricted by both human and mouse, the cellular antigenic regions of the human papillomavirus type 16 genes, containing L1   Species: Virus - HPV Sample Types: Recombinant Protein Applications: Bioassay Uncoupling of p97 ATPase activity has a dominant negative effect on protein extraction Authors: HB Rycenga, KB Wolfe, ES Yeh, DT Long Sci Rep, ;9(1)