Acceleration of vaccine development by improvement of process understanding - Analysis of the host cell proteome

While regulatory agencies require stringent product quality and safety to be upheld in biopharmaceutical products, today’s competitive biopharmaceutical market requires short process development times. The demand to accelerate especially the development of vaccines became obvious with the COVID-19 pandemic. By expanding process understanding with the use of process design tools the development time of the purification could be significantly shortened. High throughput experimentation (HTE) provides an automated experimentation platform, which minimizes the amount of used samples and saves experimental time. In this approach, HTE is used to acquire experimental data to regress parameters used as inputs for a chromatographic mechanistic model with the objective to establish an E. coli vaccine purification process development platform for a recombinant subunit vaccine. To provide a generic process development strategy that can be applied to novel antigens, the focus lies on the description of the adsorption behavior of the impurities such as host cell proteins (HCPs) during the capture step. Therefore our approach focuses on the present impurities, in specific the HCPs (Figure 1). When using the same E.coli strain the knowledge regarding the host cell proteins could be transferred to a new product. The first step is the identification of HCPs. Over a thousand HCPs are identified in the E.coli harvest sample investigated by means of mass spectrometry based proteomics. A database containing the properties of these proteins can provide assistance in the decision on chromatography resins suited for the purification process of a new developed antigen. Please click Download on the upper right corner to see the full abstract

Liderazgo y Compromiso Organizacional de los Colaboradores - Unidad Ejecutora Inversión Pública SUNAT, 2017.

El presente estudio de investigación ha sido realizado con el objetivo de determinar la influencia del liderazgo y el compromiso organizacional de los colaboradoresde la Unidad Ejecutora Inversión Pública – SUNAT, 2017y validar la hipótesis de que el liderazgo influye significativamente en el compromiso organizacional de los colaboradoresde la Unidad Ejecutora Inversión Pública – SUNAT, 2017, el problema de ejercer un liderazgo autoritario dificultan a los directivos tener una buena comunicación con los trabajadores que influya en una convivencia democrática, comprometida y participativa con el desarrollo, objetivos y los fines de la entidad. El tipo de estudio que se aplicó para el presente trabajo fue el no experimental, siendo el diseño correlacional de corte transversal, así mismo los métodos de investigación aplicados fueron el deductivo e inductivo.Se trabajó con una muestra de 20 trabajadoresde laUnidad Ejecutora Inversión Pública – SUNAT, aplicándose la técnica de la encuesta y como instrumento dos cuestionarios confiables, debidamente validados para la recolección de datos de las variables en estudio. El instrumento fue validado por expertos en gestión pública y con el coeficiente de alfa de Cronbach que estableció una confiabilidad de,894para el cuestionario de liderazgo y ,714parala de compromiso organizacional de los colaboradores de laUnidad Ejecutora Inversión Pública – SUNAT, 2017. Se procesó la información y aplicó las pruebas estadísticas correspondientes, siendo presentados en tablas y gráficos, obteniendo como resultadoentre la variable Liderazgo y Compromiso organizacional de los colaboradores de la Unidad Ejecutora Inversión Pública – SUNAT, 2017, existe una correlación directa o positiva altamente significativa dado que el coeficiente de contingencia del estadístico de prueba Sperman es R=, 923, con un sig. (Bilateral) ,000 por lo que se acepta la hipótesis de investigación y se rechaza la hipótesis nula. Que el liderazgo autoritario es el que más influye en el compromiso organizacional de los colaboradores de la Unidad Ejecutora Inversión Pública – SUNAT, 2017, dado a que 12 de los 20 encuestados consideran que se ubica en el nivel alto con 60% seguido del nivel medio con un 25% determinado por 5 encuestados y del nivel muy alto con un 15% determinado por 3 encuestados, hecho que se valida con la aplicación de la prueba de Sperman que le da un R=, 595 y un sig (bilateral),006. Estableciendo una relación directa y significativ

Model-based process development for complex vaccine mixtures

The regulations, safety and purity demands are extremely high for vaccine processes and likewise reflected in process development time and cost. Reducing time-to-market is key for pharmaceutical companies, hence saving lives and money, and therefore the need raised for systematic, general and efficient process development strategies (Hanke & Ottens, 2014). Despite the tremendous variation between vaccine purification processes, platform processes for similar types of vaccines could aid to generally accelerate the process development and would be beneficial in terms of knowledge, resources, costs and regulatory aspect. High throughput process development (HTPD) approaches can be used to establish platform processes. HTPD combines high throughput technologies and statistical or mechanistic modeling in an efficient manner. In particular mechanistic models, that aim to describe the real process based upon physical processes occurring, can be of great merit to extend the level of process understanding and thereby support in making decision regarding the process design (Pirrung et al., 2019). Please click Download on the upper right corner to see the full abstract

Repellency activity of Commiphora swynnertonii exudates against Rhipicephalus appendiculatus larvae

oai:dspace.nm-aist.ac.tz:20.500.12479/2476A research article was published by Journal of Biodiversity and Environmental Sciences (JBES) Vol. 11, 2017Evaluating plant extracts for anti-tick properties is essential towards development of alternative method for controlling tick infestation and other harmful insects. This study aimed to evaluate repellency activity of Commiphora swynnertonii hexane and chloroform extracts against Rhipicephullus appendiculatus larvae using climbing bioassay method. Concentrations of 110mg/ml, 100 mg/ml, 90mg/ml, 80mg/ml, 70mg/ml, 60mg/ml and 50mg/ml of hexane and chloroform extracts were used. C. swynnertonii chloroform and hexane extract had repellency activity in all concentrations at all-time intervals. The repellency activity was appeared to be concentration and time dependent. The results from this study strengthen the view that C. swynnertonii is the potential source of anti-tick agents

Spatial transcriptomics reveals asymmetric cellular responses to injury in the regenerating spiny mouse (Acomys) ear

In contrast to other mammals, the spiny mouse (Acomys) regenerates skin and ear tissue, which includes hair follicles, glands, and cartilage, in a scar-free manner. Ear punch regeneration is asymmetric with only the proximal wound side participating in regeneration. Here, we show that cues originating from the proximal side are required for normal regeneration and use spatially resolved transcriptomics (tomo-seq) to understand the molecular and cellular events underlying this process. Analyzing gene expression across the ear and comparing expression modules between proximal and distal wound sides, we identify asymmetric gene expression patterns and pinpoint regenerative processes in space and time. Moreover, using a comparative approach with nonregenerative rodents (Mus, Meriones), we strengthen a hypothesis in which particularities in the injury-induced immune response may be one of the crucial determinants for why spiny mice regenerate whereas their relatives do not. Our data are available in SpinyMine, an easy-to-use and expandable web-based tool for exploring Acomys regeneration-associated gene expression.Stem cells & developmental biolog

Evaluation of tick repellency activity and toxicity of commiphora swynnertonii (burtt) exudates

A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Master’s in Life Science of the Nelson Mandela African Institution of Science and TechnologyThis thesis reports the repellency activity and toxicity of Commiphora swynnertonii exudates. The exudates were extracted using hexane and chloroform solvents. The crude extracts were tested for repellency activity at concentration of 110, 100, 90, 80, 70, 60 and 50 mg/ml against Rhipicephalus appendiculatus larvae. The crude extracts were further evaluated for the acute and subacute toxicity using albino mice and rats, respectively. The results showed that repellency activity appeared to be concentration and time dependent. In acute toxicity, oral administration of C. swynnertonii hexane and chloroform extract at a doses of 500, 1000, and 2000 mg/kg body weight induced no clinical signs of toxicity in the mice during 14 days of experimental period. However, C. swynnertonii chloroform extract showed some toxic properties in some of blood parameters and in vital organs (liver and kidney) of treated mice. In subacute toxicity, C. swynnertonii hexane extract showed significant change in some of hematological and biochemical parameters. Even though rats treated with C. swynnertonii chloroform extract did not develop any observable sign of toxicity, some toxic properties was observed in some of blood parameters and in vital organs (liver and kidney). It is concluded that, C. swynnertonii hexane and chloroform extracts considered as good repellency agents against Rhipicephalus appendiculatus larvae. For the toxicity evaluation, C. swynnertonii hexane extracts was nontoxic while C. swynnertonii chloroform extract had toxic properties deduced from change in blood parameters and internal organs of treated animals

The ras-related ypt protein is an ubiquitous eukaryotic protein: isolation and sequence analysis of mouse cDNA clones highly homologous to the yeast YPT1 gene.

The YPT1 gene of the yeast Saccharomyces cerevisiae codes for a guanine nucleotide-binding protein which is essential for cell viability. Using as hybridization probe cloned yeast YPT1 gene sequences, we have isolated from cDNA libraries prepared from RNA of mouse F9 and C3H10T1/2 cells several overlapping cDNA clones with identical sequence in the regions of overlap. The cDNAs were derived from a gene, designated ypt1, which codes for a protein of 205 amino acids with 71% homology to the yeast YPT1 gene product. Amino acid sequences typical for guanine nucleotide-binding proteins and characteristic for ypt proteins are perfectly conserved in the mouse ypt1 protein. Two mRNAs of 1600 and 3200 nucleotides, originating from the mouse ypt1 gene and differing in the length of their 3'-non-translated region, were identified in mouse F9 cells and in all mouse tissues examined. A monoclonal antibody specifically recognizing the 23.5-kd yeast YPT1 protein cross-reacted with a protein of identical size on protein blots of mouse, rat, pig, bovine and human cell lines