A Comparative Investigation of Release Kinetics of Paclitaxel from Natural Protein and Macromolecular Nanocarriers in Nanoscale Drug Delivery Systems

H5N1 highly pathogenic avian influenza virus (HPAIV) infections pose a significant threat to human health, with a mortality rate of around 50%. Limited global approval of H5N1 HPAIV vaccines, excluding China, prompted the need to address safety concerns related to MDCK cell tumorigenicity. Our objective was to improve vaccine safety by minimizing residual DNA and host cell protein (HCP). We developed a downstream processing method for the cell-based H5N1 HPAIV vaccine, employing CaptoTM Core 700, a multimodal resin, for polishing. Hydrophobic-interaction chromatography (HIC) with polypropylene glycol as a functional group facilitated the reversible binding of virus particles for capture. Following the two-step chromatographic process, virus recovery reached 68.16%. Additionally, HCP and DNA levels were reduced to 2112.60 ng/mL and 6.4 ng/mL, respectively. Western blot, high–performance liquid chromatography (HPLC), and transmission electron microscopy (TEM) confirmed the presence of the required antigen with a spherical shape and appropriate particle size. Overall, our presented two-step downstream process demonstrates potential as an efficient and cost-effective platform technology for cell-based influenza (H5N1 HPAIV) vaccines.

Enhanced Downstream Processing for a Cell-Based Avian Influenza (H5N1) Vaccine

pdf/green-fluorescent-protein-discover-structure-purification-wy7lpdnjxa.pdf

Green Fluorescent Protein: discover, structure, purification, applications, and future prospects

Merozoites utilize sialic acids on the red blood cell (RBC) cell surface to rapidly adhere to and invade the RBCs. Newcastle disease virus (NDV) displays a strong affinity toward membrane-bound sialic acids. Incubation of NDV with the malaria parasites dose-dependently reduces its cellular viability. The antiplasmodial activity of NDV is specific, as incubation with Japanese encephalitis virus, duck enteritis virus, infectious bronchitis virus, and influenza virus did not affect the parasite propagation. Interestingly, NDV is reducing more than 80% invasion when RBCs are pretreated with the virus. Removal of the RBC surface proteins or the NDV coat proteins results in disruption of the virus binding to RBC. It suggests the involvement of specific protein: ligand interaction in virus binding. We established that the virus engages with the parasitized RBCs (PRBCs) through its hemagglutinin neuraminidase (HN) protein by recognizing sialic acid-containing glycoproteins on the cell surface. Blocking of the HN protein with free sialic acid or anti-HN antibodies abolished the virus binding as well as its ability to reduce parasite growth. Interestingly, the purified HN from the virus alone could inhibit the parasite's growth in a dose-dependent manner. NDV binds strongly to knobless murine parasite strain Plasmodium yoelii and restricted the parasite growth in mice. Furthermore, the virus was found to preferentially target the PRBCs compared to normal erythrocytes. Immunolocalization studies reveal that NDV is localized on the plasma membrane as well as weakly inside the PRBC. NDV causes neither any infection nor aggregation of the human RBCs. Our findings suggest that NDV is a potential candidate for developing targeted drug delivery platforms for the Plasmodium-infected RBCs.

NDV Targets the Invasion Pathway in Malaria Parasite Through Cell Surface Sialic Acid Interaction.

В статье отражены результаты теоретических исследований по изучению экологической и молекулярно-генетической характеристики вируса болезни Ньюкасла, особенностей протекания инфекции.Болезнь Ньюкасла представляет собой системную инфекцию птиц, вирус вызывает высокую заболеваемость и смертность, классифицируется как вирулентный штамм птичьего ортоавулавируса 1 (AOAV-1), принадлежащий к роду Orthoavulavirus.Спектр заболеваний у птиц делится на два широких фенотипа -авирулентная (лентогенная, легко и бессимптомно протекающее заболевание у диких водоплавающих птиц) и вирулентная (мезогенная, вызывающая респираторные или неврологические симптомы с низкой смертностью; велогенная, тяжело протекающее заболевание у домашних птиц с высокой смертностью) инфекции.Вирус болезни Ньюкасла распространяется инфицированными птицами в течение 1-2 недель после заражения с фекалиями и выделениями из дыхательных путей.Вирус высоко стабилен вне хозяина, выживает в зависимости от сезона от 7 до 30 дней.Передача NDV происходит воздушно-капельным или пероральным путем.Симптомы проявляются на 2-15 сутки после заражения.Геном вируса представлен несегментированной оц 

pdf/ecological-and-molecular-genetic-characterization-of-gkpxbffgxi.pdf

Ecological and molecular genetic characterization of newcastle disease virus

NDV belongs to the avian orthoavulavirus 1 species, genus Orthoavulavirus, family Paramyxoviridae and is responsible for Newcastle disease in poultry and other avian species. It has shown significant potential as an oncolytic virus and as a vector for vaccine delivery. NDV from infected biological sera is usually being isolated or purified with density gradient ultracentrifugation. However, it has various drawbacks, including the fact that it is time-consuming and has a restricted sample size that can be processed at one time. In our study, native agarose gel electrophoresis and DLS analysis showed that NDV carried a net negative surface charge. Thus, we purified the virus using a HiTrap QFF anion exchange column with salt elution. HA and plaque assay showed that the procedure yielded NDV particles of high purity with a recovery of over 80% with the added advantages of being a fast and straightforward operation. The purity of the virus was confirmed with SDS-PAGE and western blot analysis. The hydrodynamic volume and "dry state" diameter of the purified NDV was analyzed with the help of DLS and TEM and found to be in the range of 200-300 nm. The viruses did not exhibit any deviation from their known physical properties. The genome of the virus was also detected by amplifying a 423 bp region with RT-PCR. Our study confirmed that NDV could be effectively purified with an anion exchange column. Additionally, the procedure could be easily upscaled or downscaled based on the experimental requirements.

Isolation and characterization of NDV from biological fluids through column chromatography.

All living cells are dynamic systems capable of responding to internal and external stimuli or signals. The majority of cell signaling pathways are cascades of biomolecular reactions where the receptor, upon binding with its ligand, often undergoes a conformational change. The ligand-bound receptor then activates cascades of intracellular protein networks that help the signal to be internalized. Depending on the signal received, the cells may reorganize their metabolic activities, divide, migrate, or even undergo apoptosis. The functions of these signaling proteins can be studied with the help of classical biochemical approaches, such as western blotting, pull-down assays, and immunoprecipitation. They are often combined with modern platforms, enabling the study of these signaling molecules directly in live cells or analyze hundreds and thousands of proteins and their posttranslational modifications simultaneously. In this chapter, some of the classical signaling pathways are described, followed by the experimental approaches routinely used to study these pathways. 

Approaches and methods to study cell signaling: Linguistics of cellular communication

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Isolation and characterization of NDV from biological fluids through column chromatography.

Approaches and methods to study cell signaling: Linguistics of cellular communication