Category: <span>Uncategorized</span>

Glycosylation drives protein quality and therapeutic benefits in protein-based therapies. Recently, there has been a push in the pharmaceutical industry to improve the consistency and quality of the glycan patterns on therapeutic proteins like monoclonal antibodies. Post-translational modification begins in the endoplasmic reticulum but is finished in the Golgi where more complex glycans are added. In this study, the addition of lipids via a novel mechanism provided by the medium supplement, Cell-Ess®, improves the consistency in glycan patterns so that they are more reproducible between product batches. The effect of media supplementation with Cell-Ess on the variation of glycan patterns was measured in two different media formulations across two separate experiments. Supplementation with Cell-Ess resulted in a statistically significant reduction in the variation of glycoforms when measured by the standard error of the mean. In addition, to improved consistency, there were increased higher glycoforms or galactosylation. There was also significantly more total galactosylation and significantly fewer lower glycoforms for antibodies produced by CHO cells supplemented with Cell-Ess. These data taken together suggest that the addition of lipids via Cell-Ess results in a more functional Golgi and an associated improvement of protein quality and consistency…

Biologics Production Uncategorized

In recent years, cell therapy has been suggested as a promising approach for repair and regeneration of damaged tissues. VesCell™, a blood-derived autologous cell therapy product consisting of ex vivo enriched angiogenic cell precursors (ACPs) was developed by TheraVitae for the treatment of severe heart diseases. A non-mobilized, blood-derived cell population consisting of low density cells, termed synergetic cell population (SCP), was isolated and cultured in the presence of serum-free medium (X-Vivo 15, Lonza, Walkersville, MD, USA) supplemented with growth factors and autologous serum to yield VesCell. Significant cell numbers (>50×106) exhibiting morphological, immunocytochemical, and functional characteristics of the angiogenic cell lineage were obtained from blood samples. The ACPs expressed the hematopoietic stem cell (HSC) markers CD34, CD133 and CD117, as well as specific angiogenic markers such as vascular endothelial growth factor receptor 2 (VEGFR2) (receptor 2 [R2] is also known as kinase domain region [KDR]), CD144, and CD31…

Cell & Gene Therapy Manufacturing Uncategorized

The Vmax™ technique has been used extensively to estimate filter area requirements for normal flow filtration (NFF) processes in biopharmaceutical applications. The benefits that this technique presents over conventional flow decay methods are the speed of testing, reduced volume requirements for evaluation, and competitive testing of varying filter types/sizes — all of which present an optimized filter screening strategy and preliminary estimate of optimized filter size requirements. Filter size or filtration area requirements derived using the Vmax technique consist of contributions from both capacity and flow-time aspects of the filtration process. This article examines the relative contributions of these terms to overall filter sizing vis-à-vis the ease of fluid filterability…

Biologics Production Uncategorized

Currently, the U.S. Food and Drug Administration (FDA) recommends DNA sequencing for the structural characterization of gene transfer viral vectors in investigational New Drug Applications (INDs). While FDA provides guidelines on what should be sequenced in regard to these vectors, it provides little insight — beyond GLP/GMP (GxP) guidelines — into how the finalized sequence data should be obtained. There is presently no provision for determining which sequencing methodology (or methodologies) is most appropriate for obtaining a completed sequence characterization for each of the different vector classes, and there is no standard that outlines what DNA sequencing-specific criteria the data and data collection processes should meet in order to guarantee that the sequence is 99.99% accurate…

Nucleic Acids Research Uncategorized Viral Vectors

Apoptosis is an essential biological process that has been conserved among eukaryotic organisms throughout evolution. Apoptosis, or programmed cell death, is necessary for embryological development, tissue homeostasis, immune system maintenance and development, and as a defense mechanism against the progression of cancer and viral infection. With the advent of biotechnology and the development of associated molecular biology techniques such as recombinant DNA technology and mammalian cell culture, tissues can be extracted from organisms and have their cells cultured as single cell suspensions or adherent monolayers. Therefore, these cultures can function as living production facilities for antibodies, recombinant glycoproteins, vaccines, hormones, growth factors, and more. However, the cell’s ability to control its own death is not lost upon its manipulation from the organism to culture. Consequently, apoptosis, which is so fundamentally important in-vivo, becomes a detriment to biochemical manufacturers in-vitro…

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