scholarly journals Extended Polysaccharide Analysis within the Liposomal Encapsulation of Polysaccharides System

Materials ◽  
2020 ◽  
Vol 13 (15) ◽  
pp. 3320
Author(s):  
Roozbeh Nayerhoda ◽  
Dongwon Park ◽  
Charles Jones ◽  
Elsa N. Bou Ghanem ◽  
Blaine A. Pfeifer
Keyword(s):  
Pneumococcal Disease ◽  
Encapsulation Efficiency ◽  
Surface Protein ◽  
Protein Surface ◽  
Surface Attachment ◽  
Antigen Vaccine ◽  
Model Protein ◽  
Vaccine Carrier ◽  
Future Utility ◽  
Protein Attachment

The Liposomal Encapsulation of Polysaccharides (LEPS) dual antigen vaccine carrier system was assessed across two distinct polysaccharides for encapsulation efficiency, subsequent liposomal surface adornment with protein, adjuvant addition, and size and charge metrics. The polysaccharides derive from two different serotypes of Streptococcus pneumoniae and have traditionally served as the active ingredients of vaccines against pneumococcal disease. The LEPS system was designed to mimic glycoconjugate vaccines that covalently couple polysaccharides to protein carriers; however, the LEPS system uses a noncovalent co-localization mechanism through protein liposomal surface attachment. In an effort to more thoroughly characterize the LEPS system across individual vaccine components and thus support broader future utility, polysaccharides from S. pneumoniae serotypes 3 and 4 were systematically compared within the LEPS framework both pre- and post-surface protein attachment. For both polysaccharides, ≥85% encapsulation efficiency was achieved prior to protein surface attachment. Upon protein attachment with either a model protein (GFP) or a pneumococcal disease antigen (PncO), polysaccharide encapsulation was maintained at ≥61% encapsulation efficiency. Final LEPS carriers were also evaluated with and without alum as an included adjuvant, with encapsulation efficiency maintained at ≥30%, while protein surface attachment efficiency was maintained at ≥~50%. Finally, similar trends and distributions were observed across the different polysaccharides when assessed for liposomal zeta potential and size.

scholarly journals Design Variation of a Dual-Antigen Liposomal Vaccine Carrier System

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2809 ◽  
Author(s):  
Roozbeh Nayerhoda ◽  
Andrew Hill ◽  
Marie Beitelshees ◽  
Charles Jones ◽  
Blaine Pfeifer
Keyword(s):  
Protein Localization ◽  
Surface Protein ◽  
Potential Range ◽  
Surface Attachment ◽  
Biological Affinity ◽  
Surface Localization ◽  
Design Variation ◽  
Assessment Metrics ◽  
Vaccine Carrier ◽  
Selection Of

The enclosed work focuses on the construction variables associated with a dual-antigen liposomal carrier, delivering encapsulated polysaccharides and surface-localized proteins, which served as a vaccine delivery device effective against pneumococcal disease. Here, the goal was to better characterize and compare the carrier across a range of formulation steps and assessment metrics. Specifically, the vaccine carrier was subjected to new methods of liposomal formation, including alterations to the base components used for subsequent macromolecule encapsulation and surface attachment, with characterization spanning polysaccharide encapsulation, liposomal size and charge, and surface protein localization. Results demonstrate variations across the liposomal constructs comprised two means of surface-localizing proteins (either via metal or biological affinity). In general, final liposomal constructs demonstrated a size and zeta potential range of approximately 50 to 600 nm and −4 to −41 mV, respectively, while demonstrating at least 60% polysaccharide encapsulation efficiency and 60% protein surface localization for top-performing liposomal carrier constructs. The results, thus, indicate that multiple formulations could serve in support of vaccination studies, and that the selection of a suitable final delivery system would be dictated by preferences or requirements linked to target antigens and/or regulatory demands.

Lysozyme Release from Lipid-Based Implants

2014 ◽  
Vol 1060 ◽  
pp. 83-86
Author(s):  
Joana Portugal Mota ◽  
Alexandre Campos ◽  
Duangratana Shuwisitkul ◽  
Nuno Saraiva ◽  
Marisa Nicolai
Keyword(s):  
Controlled Release ◽  
Drug Release ◽  
Smooth Surface ◽  
Surface Protein ◽  
Protein Release ◽  
Protein Loading ◽  
Loading Efficiency ◽  
Model Protein ◽  
Preparation Techniques ◽  
Potential Use

To better understand lipid-based implants applied to proteins, lysozyme was used as a model protein and was loaded into lipid-based implants in different contents. Cylindrical and spherical implants loaded with lysozyme presented a smooth surface. Protein loading efficiency decreased with increasing lysozyme loading. Lysozyme kept its activity after different implant preparation techniques. Drug release tremendously increased by increasing protein loading from 8.9 to 13.7% (w/w), for cylindrical implants. Lysozyme release was faster from cylindrical implants when compared with the spherical ones. Furthermore, by increasing the hydrophobicity of the lipid, protein release decreased. This study demonstrates the potential use of lipid-based implants for the controlled release of proteins.

scholarly journals Mutations within the Piga gene in patients with paroxysmal nocturnal hemoglobinuria

Blood ◽  
1994 ◽  
Vol 83 (9) ◽  
pp. 2418-2422 ◽  
Author(s):  
RE Ware ◽  
WF Rosse ◽  
TA Howard
Keyword(s):  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Clinical Manifestations ◽  
Surface Protein ◽  
Convincing Evidence ◽  
Pcr Analysis ◽  
Single Nucleotide ◽  
Class A ◽  
Hematologic Disorder ◽  
Polymerase Chain ◽  
Protein Attachment

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal hematologic disorder with multiple and varied clinical manifestations. The biochemical defect in PNH resides in the incomplete enzymatic assembly of glycosylphosphatidylinositol (GPI) anchors used for surface protein attachment. In all patients tested thus far, the defect is at the level of N-acetylglucosamine attachment to phosphatidylinositol (complementation class A defect). A human cDNA, Piga, that repairs cell lines with the class A defect has been recently cloned, making Piga a candidate gene for PNH. In the current study, using highly purified GPI- deficient granulocytes, we have performed Northern blot and reverse transcriptase polymerase chain reaction (RT-PCR) analysis of Piga in four patients with PNH. In each case, we have identified a mutation in the Piga coding sequence: three frameshift mutations were found, and a single nucleotide substitution (missense) mutation was identified. Our results provide convincing evidence that alterations in the Piga gene are responsible for PNH.

scholarly journals Bacillus Subtilis Spores as Delivery System for Nasal Plasmodium Falciparum Circumsporozoite Surface Protein Immunization in a Murine Model

2021 ◽  
Author(s):  
Maria Edilene M. de Almeida ◽  
Késsia Caroline Souza Alves ◽  
Maria Gabriella Santos de Vasconcelos ◽  
Thiago Serrão Pinto ◽  
Juliane Corrêa Glória ◽  
...  
Keyword(s):  
Immune Response ◽  
Public Health Problem ◽  
Surface Protein ◽  
Humoral Response ◽  
Serum Samples ◽  
Th2 Immune Response ◽  
The World ◽  
Protein Immunization ◽  
Vaccine Carrier ◽  
Full Protection

Abstract Malaria remains a widespread public health problem in tropical and subtropical regions around the world, and there is still no vaccine available for full protection. In recent years, it has been observed that spores of Bacillus subtillis can act as a vaccine carrier and adjuvant, promoting an elevated humoral response after co-administration with antigens either coupled or integrated to their surface. In our study, B. subtillis spores from the KO7 strain were used to couple the recombinant CSP protein of P. falciparum (rPfCSP), and the nasal humoral-induced immune response in Balb/C mice was evaluated. Our results demonstrate that the spores coupled to rPfCSP increase the immunogenicity of the antigen, which induces high levels of serum IgG, and with balanced Th1/Th2 immune response, being detected antibodies in serum samples for 250 days. Therefore, the use of B. subtilis spores appears to be promising for use as an adjuvant in a vaccine formulation.

Preparation and Characterization of Copolymeric Polymersomes for Protein Delivery

Drug Research ◽  
2017 ◽  
Vol 67 (08) ◽  
pp. 458-465 ◽  
Author(s):  
Alireza Nomani ◽  
Hamed Nosrati ◽  
Hamidreza Manjili ◽  
Leila Khesalpour ◽  
Hossein Danafar
Keyword(s):  
Drug Delivery ◽  
Protein Delivery ◽  
Encapsulation Efficiency ◽  
Controlled Drug Delivery ◽  
Protein Polymer ◽  
Biodegradable Copolymers ◽  
Model Protein ◽  
Sustained Protein Release ◽  
Polymer Ratio

AbstractBiodegradable copolymeric polymersomes have been used for controlled drug delivery of proteins. These polymersomes important areas to overcome formulation associated problems of the proteins. The aim of this study was to develop polymersomes using biodegradable copolymers for delivery of bovine serum albumin (BSA) as a model protein. Encapsulated BSA by mPEG-PCL polymersomes led to formation of BSA-loaded mPEG-PCL polymersomes. The polymersomes synthesized with the protein-polymer ratio of 1:4 at 15 000 rpm gave maximum loading, minimum polydispersion with maximally sustained protein release pattern, among the prepared polymersomes. Investigation on FTIR and DSC results revealed that such a high encapsulation efficiency is due to strong interaction between BSA and the copolymer.The particles size and their morphology of polymersomes were determined by DLS and AFM.The encapsulation efficiency of BSA was 91.02%. The results of AFM showed that the polymersomes had spherical shapes with size of 49 nm.The sizes of BSA-loaded polymersomes ranged from 66.06 nm to 84.97 nm. The results showed that polymersomes exhibited a triphasic release, for BSA. Overall, the results indicated that mPEG–PCL polymersomes can be considered as a promising carrier for proteins.

scholarly journals Hydration Dynamics Near a Model Protein Surface

2004 ◽  
Vol 86 (3) ◽  
pp. 1852-1862 ◽  
Author(s):  
Daniela Russo ◽  
Greg Hura ◽  
Teresa Head-Gordon
Keyword(s):  
Protein Surface ◽  
Model Protein ◽  
Hydration Dynamics

scholarly journals PspA Family Distribution, unlike Capsular Serotype, Remains Unaltered following Introduction of the Heptavalent Pneumococcal Conjugate Vaccine

2012 ◽  
Vol 19 (6) ◽  
pp. 891-896 ◽  
Author(s):  
Christina M. Croney ◽  
Mamie T. Coats ◽  
Moon H. Nahm ◽  
David E. Briles ◽  
Marilyn J. Crain
Keyword(s):  
Pneumococcal Disease ◽  
Protein A ◽  
Surface Protein ◽  
The United States ◽  
Pediatric Hospital ◽  
Protein Antigens ◽  
Conjugate Vaccines ◽  
Reactive Protein ◽  
Serotype Replacement ◽  
Heptavalent Pneumococcal Conjugate Vaccine

ABSTRACTPneumococcal conjugate vaccines (PCVs) are recommended for the prevention of invasive pneumococcal disease (IPD) in young children. Since the introduction of the heptavalent pneumococcal vaccine (PCV7) in 2000, IPD caused by serotypes in the vaccine has almost been eliminated, and previously uncommon capsular serotypes now cause most cases of pediatric IPD in the United States. One way to protect against these strains would be to add cross-reactive protein antigens to new vaccines. One such protein is pneumococcal surface protein A (PspA). Prior to 2000, PspA families 1 and 2 were expressed by 94% of isolates. Because PCV7 vaccine pressure has resulted in IPD caused by capsular serotypes that were previously uncommon and unstudied for PspA expression, it was possible that many of the new strains expressed different PspA antigens or even lacked PspA. Of 157 pediatric invasive pneumococcal isolates collected at a large pediatric hospital in Alabama between 2002 and 2010, only 60.5% had capsular serotypes included in PCV13, which came into general use in Alabama after our strains were collected. These isolates included 17 serotypes that were not covered by PCV13. Nonetheless, pneumococcal capsular serotype replacement was not associated with changes in PspA expression; 96% of strains in this collection expressed PspA family 1 or 2. Continued surveillance will be critical to vaccine strategies to further reduce IPD.

scholarly journals Immunization with Pneumococcal Surface Protein K of Nonencapsulated Streptococcus pneumoniae Provides Protection in a Mouse Model of Colonization

2015 ◽  
Vol 22 (11) ◽  
pp. 1146-1153 ◽  
Author(s):  
Lance E. Keller ◽  
Xiao Luo ◽  
Justin A. Thornton ◽  
Keun-Seok Seo ◽  
Bo Youn Moon ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Mouse Model ◽  
Pneumococcal Disease ◽  
Surface Protein ◽  
Antibody Isotype ◽  
Content Type ◽  
Mouse Immunization ◽  
Escape Mutants

ABSTRACTCurrent vaccinations are effective against encapsulated strains ofStreptococcus pneumoniae, but they do not protect against nonencapsulatedStreptococcus pneumoniae(NESp), which is increasing in colonization and incidence of pneumococcal disease. Vaccination with pneumococcal proteins has been assessed for its ability to protect against pneumococcal disease, but several of these proteins are not expressed by NESp. Pneumococcal surface protein K (PspK), an NESp virulence factor, has not been assessed for immunogenic potential or host modulatory effects. Mammalian cytokine expression was determined in anin vivomouse model and in anin vitrocell culture system. Systemic and mucosal mouse immunization studies were performed to determine the immunogenic potential of PspK. Murine serum and saliva were collected to quantitate specific antibody isotype responses and the ability of antibody and various proteins to inhibit epithelial cell adhesion. Host cytokine response was not reduced by PspK. NESp was able to colonize the mouse nasopharynx as effectively as encapsulated pneumococci. Systemic and mucosal immunization provided protection from colonization by PspK-positive (PspK+) NESp. Anti-PspK antibodies were recovered from immunized mice and significantly reduced the ability of NESp to adhere to human epithelial cells. A protein-based pneumococcal vaccine is needed to provide broad protection against encapsulated and nonencapsulated pneumococci in an era of increasing antibiotic resistance and vaccine escape mutants. We demonstrate that PspK may serve as an NESp target for next-generation pneumococcal vaccines. Immunization with PspK protected against pneumococcal colonization, which is requisite for pneumococcal disease.

scholarly journals Evidence for Vaccine Synergy between Borrelia burgdorferi Decorin Binding Protein A and Outer Surface Protein A in the Mouse Model of Lyme Borreliosis

2000 ◽  
Vol 68 (11) ◽  
pp. 6457-6460 ◽  
Author(s):  
Mark S. Hanson ◽  
Nita K. Patel ◽  
David R. Cassatt ◽  
Nancy D. Ulbrandt
Keyword(s):  
Borrelia Burgdorferi ◽  
Outer Surface ◽  
Binding Protein ◽  
Protein A ◽  
Surface Protein ◽  
Outer Surface Protein A ◽  
Outer Surface Protein ◽  
Antigen Vaccine ◽  
Single Antigen

ABSTRACT Mice immunized with either the predominantly vector-stage lipoprotein outer surface protein A (OspA) or the in vivo-expressed lipoprotein decorin binding protein A (DbpA) are protected againstBorrelia burgdorferi challenge. DbpA-OspA combinations protected against 100-fold-higher challenge doses than did either single-antigen vaccine and conferred significant protection against heterologous B. burgdorferi, B. garinii, andB. afzelii isolates, suggesting that there is synergy between these two immunogens.

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