Efficient cellular and humoral immune response and production of virus-neutralizing antibodies by the Hepatitis B Virus S/preS1 16-42 antigen

Despite the availability of improved antiviral therapies, infection with Hepatitis B virus (HBV) remains a3 significant health issue, as a curable treatment is yet to be discovered. Current HBV vaccines relaying on the efficient expression of the small (S) envelope protein in yeast and the implementation of mass vaccination programs have clearly contributed to containment of the disease. However, the lack of an efficient immune response in up to 10% of vaccinated adults, the controversies regarding the seroprotection persistence in vaccine responders and the emergence of vaccine escape virus mutations urge for the development of better HBV immunogens. Due to the critical role played by the preS1 domain of the large (L) envelope protein in HBV infection and its ability to trigger virus neutralizing antibodies, including this protein in novel vaccine formulations has been considered a promising strategy to overcome the limitations of S only-based vaccines. In this work we aimed to combine relevant L and S epitopes in chimeric antigens, by inserting preS1 sequences within the external antigenic loop of S, followed by production in mammalian cells and detailed analysis of their antigenic and immunogenic properties. Of the newly designed antigens, the S/preS116–42 protein assembled in subviral particles (SVP) showed the highest expression and secretion levels, therefore, it was selected for further studies in vivo. Analysis of the immune response induced in mice vaccinated with S/preS116–42- and S-SVPs, respectively, demonstrated enhanced immunogenicity of the former and its ability to activate both humoral and cellular immune responses. This combined activation resulted in production of neutralizing antibodies against both wild-type and vaccine-escape HBV variants. Our results validate the design of chimeric HBV antigens and promote the novel S/preS1 protein as a potential vaccine candidate for administration in poor-responders to current HBV vaccines.publishedVersio

Efficient cellular and humoral immune response and production of virus-neutralizing antibodies by the Hepatitis B Virus S/preS116-42 antigen

Despite the availability of improved antiviral therapies, infection with Hepatitis B virus (HBV) remains a3 significant health issue, as a curable treatment is yet to be discovered. Current HBV vaccines relaying on the efficient expression of the small (S) envelope protein in yeast and the implementation of mass vaccination programs have clearly contributed to containment of the disease. However, the lack of an efficient immune response in up to 10% of vaccinated adults, the controversies regarding the seroprotection persistence in vaccine responders and the emergence of vaccine escape virus mutations urge for the development of better HBV immunogens. Due to the critical role played by the preS1 domain of the large (L) envelope protein in HBV infection and its ability to trigger virus neutralizing antibodies, including this protein in novel vaccine formulations has been considered a promising strategy to overcome the limitations of S only-based vaccines. In this work we aimed to combine relevant L and S epitopes in chimeric antigens, by inserting preS1 sequences within the external antigenic loop of S, followed by production in mammalian cells and detailed analysis of their antigenic and immunogenic properties. Of the newly designed antigens, the S/preS116–42 protein assembled in subviral particles (SVP) showed the highest expression and secretion levels, therefore, it was selected for further studies in vivo. Analysis of the immune response induced in mice vaccinated with S/preS116–42- and S-SVPs, respectively, demonstrated enhanced immunogenicity of the former and its ability to activate both humoral and cellular immune responses. This combined activation resulted in production of neutralizing antibodies against both wild-type and vaccine-escape HBV variants. Our results validate the design of chimeric HBV antigens and promote the novel S/preS1 protein as a potential vaccine candidate for administration in poor-responders to current HBV vaccines

Optimized Technologies for Cointegration of MOS Transistor and Glucose Oxidase Enzyme on a Si-Wafer

The biosensors that work with field effect transistors as transducers and enzymes as bio-receptors are called ENFET devices. In the actual paper, a traditional MOS-FET transistor is cointegrated with a glucose oxidase enzyme, offering a glucose biosensor. The manufacturing process of the proposed ENFET is optimized in the second iteration. Above the MOS gate oxide, the enzymatic bioreceptor as the glucose oxidase is entrapped onto the nano-structured TiO2 compound. This paper proposes multiple details for cointegration between MOS devices with enzymatic biosensors. The Ti conversion into a nanostructured layer occurs by anodization. Two cross-linkers are experimentally studied for a better enzyme immobilization. The final part of the paper combines experimental data with analytical models and extracts the calibration curve of this ENFET transistor, prescribing at the same time a design methodology

One-pot synthesis of superabsorbant hybrid hydrogels based on methacrylamide gelatin and polyacrylamide: effortless control of hydrogel properties through composition design

BiocompatibLe methacryLamide-modified geLatin (GELMA) hydrogeLs with tuned characteristics, obtained through network-forming photopoLymerization, have recenty attracted increasing attention due to their wide range of possibLe appLications such as drug reLease, tissue regeneration and generation of bioartificiaL impLants. Due to the controlled number of C=C bonds, GELMA may simuLtaneousLy act as macromonomer and crossLinker Leading through poLymerization to hydrogeLs with rationally designed performances. This study provides effortess one-pot synthesis of hybrid hydrogeLs based on covaLenty Linked GELMA and poLyacryLamide (PM), using photo-induced network-forming poLymerization. ConventionaL synthesis of simiLar hydrogeLs Leads to interpenetrating geLatin and PAA networks, usually invoLving muLtistep crossLinking of the components and the use of toxic crossLinkers. Through the described one-pot chemistry, the synthetic water superabsorbent PM with its well-recognized advantages can rationally benefit from the high biocompatibiLity and cell-adherence of GELMA in a simpLe covaLent way. This work provides a correLation between the composition and the corresponding hydrogeL properties (incLuding swelling, pH influence, mechanicaL behaviour, abiLity to generate porous scaffoLds, enzymatic degradation). The addition of PM moduLated the network density and the water affinity allowing the controL of eLasticity and degradabiLity. SuppLementary crossLinking of the synthetic component provided additionaL controL over hydrophiLicity. The capacity of such hydrogeLs to generate porous scaffoLds was proved; interesting morphoLogies were deveLoped onLy by varying the composition. In vitro ceLLuLar studies indicated that the presence of GELMA conferred controlled cell-affinity properties to the bicomponent hydrogeLs. NevertheLess, the drug reLease potentiaL of such hydrogeLs was preLiminarlly investigated using sodium nafcillin. GELMA PM hydrogeLs may be usefuL for tissue regeneration due to effortess synthesis, compositionaL flexibiLity and variabLe properties

Dielectrophoretic and Electrical Impedance Differentiation of Cancerous Cells Based on Biophysical Phenotype

Here, we reported a study on the detection and electrical characterization of both cancer cell line and primary tumor cells. Dielectrophoresis (DEP) and electrical impedance spectroscopy (EIS) were jointly employed to enable the rapid and label-free differentiation of various cancer cells from normal ones. The primary tumor cells that were collected from two colorectal cancer patients, cancer cell lines (SW-403, Jurkat, and THP-1), and healthy peripheral blood mononuclear cells (PBMCs) were trapped first at the level of interdigitated microelectrodes with the help of dielectrophoresis. Correlation of the cells dielectric characteristics that was obtained via electrical impedance spectroscopy (EIS) allowed evident differentiation of the various types of cell. The differentiations were assigned to a “dielectric phenotype” based on their crossover frequencies. Finally, Randles equivalent circuit model was employed for highlighting the differences with regard to a series group of charge transport resistance and constant phase element for cancerous and normal cells

Technology transfer of oil-in-water emulsion adjuvant manufacturing for pandemic influenza vaccine production in Romania: preclinical evaluation of split virion inactivated H5N1 vaccine with adjuvant

Millions of seasonal and pandemic influenza vaccine doses containing oil-in-water emulsion adjuvant have been administered in order to enhance and broaden immune responses and to facilitate antigen sparing. Despite the enactment of a Global Action Plan for Influenza Vaccines and a multi-fold increase in production capabilities over the past 10 years, worldwide capacity for pandemic influenza vaccine production is still limited. In developing countries, where routine influenza vaccination is not fully established, additional measures are needed to ensure adequate supply of pandemic influenza vaccines without dependence on the shipment of aid from other, potentially impacted first-world countries. Adaptation of influenza vaccine and adjuvant technologies by developing country influenza vaccine manufacturers may enable antigen sparing and corresponding increases in global influenza vaccine coverage capacity. Following on previously described work involving the technology transfer of oil-in-water emulsion adjuvant manufacturing to a Romanian vaccine manufacturing institute, we herein describe the preclinical evaluation of inactivated split virion H5N1 influenza vaccine with emulsion adjuvant, including immunogenicity, protection from virus challenge, antigen sparing capacity, and safety. In parallel with the evaluation of the bioactivity of the tech-transferred adjuvant, we also describe the impact of concurrent antigen manufacturing optimization activities. Depending on the vaccine antigen source and manufacturing process, inclusion of adjuvant was shown to enhance and broaden functional antibody titers in mouse and rabbit models, promote protection from homologous virus challenge in ferrets, and facilitate antigen sparing. Besides scientific findings, the operational lessons learned are delineated in order to facilitate adaptation of adjuvant technologies by other developing country institutes to enhance global pandemic influenza preparedness