Cancer is one of the leading causes of death worldwide, and the second leading cause of death in Cuba. To address this serious health problem, some research has involved suppressing tumor growth by inhibiting the angiogenesis process using several molecules including antibodies. A divalent version of antibody fragments, the CIGB-598a, with a molecular weight between 100 and 110 kDa, has been expressed in CHO cells specific for a novel epitope of the human vascular endothelium growth factor (VEGF). This material has been generated at the Center for Genetic Engineering and Biotechnology to support cancer research efforts. As in other studies involving the purification of recombinant molecules, CIGB-598a exhibited a high degree of aggregation in the CHO cell culture supernatant. This required the design of a downstream process capable of removing high levels of aggregates to obtain a highly pure target molecule for use in preclinical studies and human applications further down the road. We have developed a suitable downstream method based on the combination of three chromatography processes: affinity, cation-exchange, and anion-exchange that recover a relatively low level of CIGB-598a, but at a level of high purity (greater than 95 %) with fewer aggregates (below 1%)…
BioProcessing Journal Posts
Cryopreservation is a desirable method for the long-term storage of human red blood cells (RBCs). Current protocols employ high concentrations of glycerol that must be removed from thawed RBCs prior to transfusion. Small-molecule ice recrystallization inhibitors (IRI) can protect RBCs from cryoinjury during the freezing and thawing process in the presence of reduced amounts of glycerol. Although reducing the concentration of glycerol during freezing reduces post-thaw deglycerolization times, thawed RBC units still require post-thaw processing. Herein, we report the cryopreservation of RBCs using the non-permeating cryoprotective agent (CPA) hydroxyethyl starch (HES) supplemented with small-molecule IRIs: (1) PMP-Glc (110 mM); and (2) pBrPh-Glc (30 mM). The results demonstrate that 30 mM pBrPh-Glc in 11.5 % (w/w) HES affords quantitative post-thaw recovery of intact RBCs that are superior to those obtained using glycerol with slow cooling rates, and show the utility of small-molecule IRIs in cryopreservation…
A rapid increase in the number of gene therapy trials and products has led to a comparable increase in the need for industrial production of viral gene therapy vectors such as lentiviral, adeno-associated, and adenoviral vectors. Current production systems are limited with respect to scalability and robustness. With our CAP® and CAP-T™ cell lines, we have developed a novel system for high-density suspension culture, efficient and reproducible transfection, and highly efficient production of viral vectors. By upstream process optimization, we have obtained a robust and high-density fed-batch culture system which can be scaled in any current bioreactor format. A design-of-experiments approach has been employed to optimize transient production of lentiviral vectors with significantly higher titers than can be obtained with adherent HEK293T cells…
One of the objectives of the upcoming ICH 12: Pharmaceutical Lifecycle Management guidance is to manage product development and manufacturing process information in order to establish and maintain appropriate change control over the entire product lifecycle. The 2014 ICH Q12 Concept Paper also stresses ICH Q12’s role in connecting ICH Q8 through ICH Q11 into a complete lifecycle approach to assure product quality and continuous improvement of manufacturing operations. However, neither the Concept Paper or subsequent public discussion and presentations appear to identify the ICH Q8 design space as a central mechanism for collecting and maintaining product and process information…
You have worked tirelessly on your project and an opportunity arises to present your results. It might be a group meeting, departmental seminar, job interview, or scientific conference. Alternatively, you may need to raise money to fund your research, start a biotechnology company, or market a new product. In each of these instances, delivering your message effectively is essential to achieving your goal and earning the respect and admiration of colleagues…
The treatment of animal serum by gamma irradiation is performed to mitigate the risk of introducing undesired microorganisms (viruses, mollicutes, or other microbes) into a cell culture. Serum manufacturers and end-users utilize irradiation contractors to perform this process. The irradiation process must be validated, which involves establishing the: (A) minimum dose that achieves the required inactivation of the microorganisms of interest; (B) maximum acceptable dose at which the serum still maintains all of its required functional specifications; and (C) process used by the contract irradiator that allows treatment of the serum product within these defined limits. In the present article, we describe the best practices for qualifying the distribution and magnitude of absorbed dose (performance qualification [PQ] dose-mapping) when serum is gamma irradiated. PQ dose-mapping includes the following: (1) documentation of dose distribution characteristics in defined product load configurations for a specified pathway through the irradiator; (2) assessment of the process capability of the defined product load configurations and irradiation pathway for respecting the dose specification for the serum; and (3) development of a method for routine dose monitoring of the irradiation process with the defined product load configurations and the specified irradiation pathway…
I have to begin with a disclosure. I am a card-carrying, old-school, hypercritical academic who has always been among the first to complain about going to a conference and having to sit through an infomercial disguised as a science talk. But, perhaps I have a disguise of my own. I actually enjoy hearing talks focused on applied research and product development at least as much as more “academic” talks. I admire and respect the practical application of brilliant science. I believe that’s what scientists are supposed to do…
Filamentous fungi are an important class of organisms with significant commercial importance in the biotechnology sector. Those that dominate the markets and are deemed the most important include: Aspergillus, Penicillium, and Trichoderma. They have become indispensable for the production of enzymes due to their exceptionally high capacity to express and secrete proteins. In addition, recent advancements in genomics have allowed for the sequencing or partial sequencing of many of these species. The significance of these fungi and their associated enzymes has accelerated interest in understanding the mechanisms involved in the regulation of gene expression, primarily through the characterisation of the transcriptional elements involved…
Biological samples such as human tissue, cell lines, and serum specimens are a direct source of molecular data that can be used to identify targets for disease therapy, detection, and prevention. However, the validity and reliability of data derived from burgeoning analytical technology is highly dependent upon properly preserved, well-annotated biospecimens. As a result, biopreservation and biobanking have become an increasingly integral component in advancing today’s biotherapeutic development…