Impact of harvest conditions on the glycosylation profile of a therapeutic antibody

It is essential to maintain the glycosylation profile of a protein therapeutic as it can affect its efficacy (ADCC or CDC), pharmacokinetics/pharmacodynamics, safety, and/or immunogenicity. Pilot scale experiments for an antibody manufacturing process, which used low pH during harvest step to improve the robustness of a subsequent downstream filtration step, revealed the presence of certain peaks that were not present in the harvested material without such low pH treatment. Analytical characterization of these peaks identified them to be associated with M6, M7, M8, and M9 high mannose glycoforms. Further experiments, which looked into the individual effects of the feed pump and continuous centrifuge in the pilot scale harvest set up, concluded that the glycoforms were enriched in high mannose forms during the continuous centrifugation step. Intriguingly, the same was not observed when the experiments were done using a bench top centrifuge. This led to the hypothesis that the cells were exposed to varying shear rates across scales. The hypothesis was subsequently tested using the pilot scale centrifuge and a capillary shear device, in which the cells were exposed to different levels of shear stress. The data from the experiments confirmed that it was indeed the combination of high shear and low pH that resulted in the enrichment of high mannose glycoforms in the centrifuged material. Results from subsequent experiments also suggested that the enrichment potentially occurred due to the release of immature glycoforms associated with the endoplasmic reticulum (ER) and Golgi apparatus

Evaluation of Satellite-Based Rainfall Estimates in the Lower Mekong River Basin (Southeast Asia)

Satellite-based precipitation is an essential tool for regional water resource applications that requires frequent observations of meteorological forcing, particularly in areas that have sparse rain gauge networks. To fully realize the utility of remotely sensed precipitation products in watershed modeling and decision-making, a thorough evaluation of the accuracy of satellite-based rainfall and regional gauge network estimates is needed. In this study, Tropical Rainfall Measuring Mission (TRMM) Multi-Satellite Precipitation Analysis (TMPA) 3B42 v.7 and Climate Hazards Group InfraRed Precipitation with Station data (CHIRPS) daily rainfall estimates were compared with daily rain gauge observations from 2000 to 2014 in the Lower Mekong River Basin (LMRB) in Southeast Asia. Monthly, seasonal, and annual comparisons were performed, which included the calculations of correlation coefficient, coefficient of determination, bias, root mean square error (RMSE), and mean absolute error (MAE). Our validation test showed TMPA to correctly detect precipitation or no-precipitation 64.9% of all days and CHIRPS 66.8% of all days, compared to daily in-situ rainfall measurements. The accuracy of the satellite-based products varied greatly between the wet and dry seasons. Both TMPA and CHIRPS showed higher correlation with in-situ data during the wet season (JuneSeptember) as compared to the dry season (NovemberJanuary). Additionally, both performed better on a monthly than an annual time-scale when compared to in-situ data. The satellite-based products showed wet biases during months that received higher cumulative precipitation. Based on a spatial correlation analysis, the average r-value of CHIRPS was much higher than TMPA across the basin. CHIRPS correlated better than TMPA at lower elevations and for monthly rainfall accumulation less than 500 mm. While both satellite-based products performed well, as compared to rain gauge measurements, the present research shows that CHIRPS might be better at representing precipitation over the LMRB than TMPA

Efficacy of frog skin lipids in wound healing

<p>Abstract</p> <p>Background</p> <p>Frog skin has been sequentially and scientifically evaluated by our group for its wound healing efficiency. Owing to the complex structure of skin, attempts were being made to analyse the role of individual constituents in different phases of healing. Our earlier papers have shown the significance of frog skin not only in wound healing but also enhancing the proliferating activity of the epidermal and dermal cells which are instrumental for normal healing process. We also have identified for the first time novel antimicrobial peptides from the skin of <it>Rana tigerina </it>and thereby reduce the complications involved in the sepsis.</p> <p>Purpose of the study and Results</p> <p>The current study envisages the role of frog skin lipids in the inflammatory phase of wound healing. The lipid moiety of the frog skin dominated by phospholipids exhibited a dose dependent acceleration of healing irrespective of the mode of application. The efficiency of the extract is attributed partially to the anti-inflammatory activity as observed by the histochemical and immunostimulatory together with plethysmographic studies.</p> <p>Conclusions</p> <p>Thus, frog skin for the first time has been demonstrated to possess lipid components with pharmaceutical and therapeutic potential. The identification and characterization of such natural healing molecules and evaluating their mechanism of action would therefore provide basis for understanding the cues of Nature and hence can be used for application in medicine.</p

Fully integrated continuous antibody processing demonstrates improved productivity

Continuous processing has been an important topic in the biopharmaceutical industry for the past few years. However, as this technology is still at an early stage, continuous processing has remained largely siloed to individual steps with little thought towards combining each stage to create a fully continuous process. This presentation will show a fully integrated continuous process, from production bioreactor to drug substance at pilot scale, as well as touch on considerations for manufacturing costs and facility throughput. Specific emphasis will be given towards the production perfusion bioreactor, capture using multicolumn chromatography, viral inactivation, and buffer exchange

TPX: Biomedical literature search made easy

TPX is a web-based PubMed search enhancement tool that enables faster article searching using an alysis and exploration features . These features include identification of relevant biomedical concepts from search results with linkouts to source databases, concept based article categorization, concept assisted search and filtering, query refinement. A distinguishing feature here is the ability to add user-defined concept names and/or concept types for named entity recognition. The tool allows contextual exploration of knowledge sources by providing concept association maps derived from the MEDLINE repository. It also has a full-text search mode that can be configured on request to access local text repositories, incorporating entity co-occurrence search at sentence/paragraph levels. Local text files can also be analyzed on-the-fly

Developing integrated platforms for the generation of cell lines expressing bispecific proteins with desired qualities

Complex bispecific and novel molecules are increasingly being developed as therapeutic proteins. These molecules present challenges for the generation of high quality manufacturing cell lines with good productivity and desirable product quality. High aggregation, poor sialylation and multiple chain mispairing are three of the major product quality problems affecting these molecules. To address these issues, we have developed a new expression platform which has been combined with high throughput analytical assays to screen and select clones with the preferred product quality profiles. Case studies will be presented on the development of cell lines expressing two types of bispecific molecules. To address the high level of aggregation observed with a bispecific molecule, the underlying cause of aggregation has been investigated and vector engineering tools have been applied to manipulate the expression of specific subunits of the protein leading to reduced aggregation. For another monomeric bispecific project, cell lines expressing four different polypeptides are generated and then efficiently screening at early stages of development to identify those with a high proportion of correctly assembled protein. These results also provide valuable information to assist in the molecular design and protein engineering of bispecific molecules

Heterogeneous microbial oceanographic environments: Application of GIS technology in deciphering of microenvironment scenarios off the central west coast of India

In the vast oceanic microbial environment of 2468.83km 2, GIS modeling techniques involving sixty query steps, enabled the deciphering of Microenvironments as low as 1.19km 2 to 38.6 km 2 for the summer of 2004 and in case of summer 2005 where 84 query steps were involved to decipher Microenvironments of 10.55km 2 to 25.94km 2. Thirtythree sampling stations were established between Betul to Ankola off the central west coast of India accounting for a spatial coverage of 2468.83km 2. GIS query-modeling investigation was carried out using spatial layers of depth, optical parameters (k-Irradiance attenuation Coefficient, c-Beam attenuation coefficient), sediment size parameters (Sediment Mean Size and Sediment Sorting) and Benthic Foraminifera Suborders (Rotaliina, Textulariina, Miliolina, Lagenina). Foraminifera have been used as a surrogate parameter. However, any microbial parameter could proxy for foraminifers providing for the numerical deciphering of microenvironments. This is suggestive of the assimilation of GIS technology for a better appreciation of microbial oceanography

Perioperative Management of Adult Patients With External Ventricular and Lumbar Drains: Guidelines From the Society for Neuroscience in Anesthesiology and Critical Care

External ventricular drains and lumbar drains are commonly used to divert cerebrospinal fluid and to measure cerebrospinal fluid pressure. Although commonly encountered in the perioperative setting and critical for the care of neurosurgical patients, there are no guidelines regarding their management in the perioperative period. To address this gap in the literature, The Society for Neuroscience in Anesthesiology & Critical Care tasked an expert group to generate evidence-based guidelines. The document generated targets clinicians involved in perioperative care of patients with indwelling external ventricular and lumbar drains