Only a year ago, the lack of standards for single-use equipment systems (SUS), as covered in this journal, was cited as inhibiting growth in the bioprocessing industry. The need to establish more standardized approaches for single-use devices’ design, quality, and leachables/extractables that regulators and industry end-users could accept was slowing the acceptance rate of these novel devices…
BioProcessing Journal Posts
Automobiles contribute significantly to CO2 emissions in the atmosphere. However, laboratory equipment such as CO2 incubators can potentially release over 120,000 liters of CO2 gas per year. The “epGreen initiative” established by Eppendorf® will ensure that their products are developed with the mindset of reducing environmental impact…
There are an estimated 225 million cases of malaria each year leading to nearly a million deaths, mostly in children under five years of age. Of the five malaria parasite species that can infect humans Plasmodium falciparum is by far the most virulent, causing 89% of all malaria related deaths in 2008. The P. falciparum parasite escapes the immune system by inserting antigens, functioning as adhesions, into the membrane of the infected erythrocyte. This allows it to sequester in the host microvasculature, thereby avoiding clearance in the spleen…
In an attempt to express and isolate BoLA-NC3*00101, a bovine, non-classical, major histocompatibility complex (MHC) class I protein, the mouse mastocytoma cell line P815 was transfected with the NC3*00101 transgene. The transfected cells were checked for expression by flow cytometry and positive stable transfectants were sorted using a fluorescence-activated cell sorter (FACS). A large amount of purified protein is required for the generation of monoclonal antibodies in order to immunize the mice and then boost the host for several months to get highly-specific antibodies. To avoid using a large number of cell culture flasks and shorten the time required to obtain a sufficient quantity of protein, stably-transfected BoLA-NC3*00101-expressing cells were grown in a pitched-blade bioreactor. One week of culturing in the bioreactor yielded a large cell mass which was used to isolate and purify the protein…
The manufacturing of cell-based therapies requires harmonized processing protocols to ensure consistent quality. Expanded adipose-derived mesenchymal stromal cells (ASC) are among the most promising candidates for such therapies due to their regenerative and immunomodulatory properties. However, transitioning these therapies to large-scale production presents challenges related to cell recovery, formulation, and in-process cell counting (IPC). The Gibco CTS™ Rotea™ multipurpose Counterflow Centrifugation System enables multiple workflow operations—including cell separation, concentration, washing, and buffer exchange—and is designed to operate within a GMP environment. From a cell concentration perspective, the Rotea system can also formulate concentrated cells in different types of media, including fetal bovine serum…
The recent decision of the US Supreme Court in Assoc. Mol. Pathol. v. Myriad Genetics, Inc., No. 12-398 (June 13, 2013) continued the Court’s efforts to clarify those innovations eligible for patent protection. Myriad unanimously held that “isolated” BRCA1/2 DNAs were patent ineligible under 35 USC § 1 01 while BRCA1/2 complementary DNAs (cDNAs) were patent eligible under that statutory provision. In considering patent eligibility, the Court proposed a test for isolated DNAs that balances incentives and impediments to innovation arising from patent protection, without placing any weight on whether the innovation relates to nature or to an abstract idea. The Court concluded that the mere act of isolating DNA segments claimed in terms of their genetic information was insufficient to render the claims patent eligible. Although the full effect of Myriad won’t be fully known for some time, it is already apparent that reliance expectations developed over 30 years have been upset. Taking a closer look, Myriad discovered the BRCA1 and BRCA2 genes and their influence on the risks of breast and ovarian cancers. Myriad also located these genes in the human genome, isolated the genes, and determined the sequences of several alleles, or versions, of these genes, with some alleles associated with higher cancer risks than others. This led Myriad to develop and market a diagnostic test for assessing breast and ovarian cancer risk, and to seek patent protection for the technology. Myriad refused to license the technology to competitors, which led to a declaratory judgment suit against Myriad. The district court granted summary judgment to the challengers, holding that Myriad’s isolated BRCA1/2 DNA claims and BRCA1/2 cDNA claims were invalid for patent ineligibility under 35 USC § 101. For reasons not relevant to the patent eligibility of DNA composition claims, the Court of Appeals for the Federal Circuit (CAFC) had two cracks at this case on appeal, and both times it held that isolated BRCA1/2 DNAs and BRCA1/2 cDNAs were patent eligible. On its own second review of the case, the US Supreme Court unanimously and finally decided the appeal of summary judgment, holding isolated BRCA1/2 DNAs patent ineligible, but BRCA1/2 cDNAs patent eligible…
Heat inactivation is dependent both on temperature and time at temperature, making inter-assay and intervirus comparisons of heat sensitivity of viruses problematic. Historically, heat inactivation data for pathogens, including viruses, have been evaluated by determining decimal reduction value ([D] the time required to inactivate 1 log10 of the organism at a given temperature) and the incremental temperature required to decrease the D by 1 log10 (z). We recommend the use of a straightforward approach for extrapolating heat inactivation (i.e., inactivation vs. time at fixed temperature) data from measured to non-measured temperatures that is based not on the z value, but on a power function fit of the D vs. temperature plots. There needs to have been at least three temperatures evaluated in the inactivation vs. time kinetics studies in order to conduct these modeling analyses. For inter-assay and inter-virus comparisons of heat inactivation sensitivity, we propose the use of two modeled parameters: (1) temperature required to inactivate 1 log10 of virus in 0.5 minutes; and (2) time required for 1 log10 reduction in infectivity at 80°C. By using both modeled parameters, we have calculated consensus heat inactivation values for two caliciviruses (feline calicivirus and murine norovirus)…
Novavax reported that, in 28 days following public release of the avian influenza A/Anhui/1/2013 (H7N9) virus gene sequences, its recombinant DNA and baculovirus-Spodoptera frugiperda cell culture-based technology was used to produce a virus-like particle vaccine to avian influenza A(H7N9) virus, and murine animal challenge studies were initiated. This report describes Novavax’s manufacturing process and the coordinated timing of critical activities necessary to produce and release a clinical batch of avian influenza A(H7N9) virus virus-like particle vaccine, under current good manufacturing practices, within three months from the time that the virus genomic sequences for this potential pandemic influenza virus were reported…