In order to unravel new protein activities and functions, we have expressed and purified a large number of human proteins. We have chosen to study secreted proteins and the extra-cellular domains of putative single transmembrane domain-containing proteins. In order to retain the natural protein characteristics as far as possible, we have used a mammalian expression system. Human embryonic kidney (HEK293) cells were chosen as they have been shown to possess a high protein-secretory potential. The secreted proteins were expressed with a carboxy-terminal tag and purified by affinity chromatography. Each protein was produced at a routine scale from 500 ml cell cultures, and the secreted protein was purified from the culture supernatant…
Category: <span>Research</span>
The recent discovery of cancer stem cells in leukemia, brain cancer, and breast cancer has had a significant impact on cancer research and how cancers are thought to arise. Their discovery has resulted in unique, but pressing challenges that may ultimately only be resolved through the development of large-scale bioprocesses. The scarcity of these cells is currently impeding discovery of new cancer treatments that specifically aim to eradicate the population of tumour cells thought to be responsible for tumour growth and metastasis. Fundamental bioprocessing principles have been applied to develop scalable, large-scale cultures for cancer stem cells, to address this issue of cancer stem cell scarcity. Development of such bioprocesses differs significantly from other, more conventional cultures since it is the cells themselves that are of interest rather than the products of the cells (e.g., proteins)…
The efficacy of pharmaceuticals, nutraceuticals, and cosmeceuticals depends not only on the chemical composition of the active ingredients, but also on the formulation of the product and the method of delivery. Nanoemulsions—formulations that encapsulate drug or nutrient molecules in sub-micron-sized oil droplets suspended in water—have been found to increase the rate and level of absorption into the body, and offer other benefits including the mitigation of side-effects as well as an antiviral and antibacterial action. With an emulsifier layer surrounding the droplet, nanoemulsions exhibit high levels of stability for extended periods of six months or more…
Many precautions are taken in a typical research lab to ensure the integrity of biological specimens. Temperature, storage, and personnel access, among others, are all tightly controlled, and codified into standard operating procedures (SOPs), if not almost biblical law. And no wonder — companies have millions invested in biotech solutions whose progress is often measured in years and decades. Scientists have their life’s work on the line. So, it is with some surprise that the diligence most companies exercise during the research and development process is not always maintained during specimen transport. Every time a specimen leaves the lab, be it for further analytical testing or investigational purposes, it runs a heightened risk of contamination, especially from fluctuating temperatures. Ensuring that this does not happen should be the responsibility and concern of everyone with a stake in a biological product’s success…
The significance of tissue culture is that an understanding of animal physiology will come from a study of the basic elements of the animal (the cell) in isolated, pure form (clonal culture), under defined conditions (hormonally defined media). This should be the fundamental approach to animal physiology for the foreseeable future. The problem with this formulation in 1958, when I began work on tissue culture, was that cells in culture did not display the differentiated properties of the tissue of origin…
The quantitative and qualitative analysis of proteins and their amino acid sequence composition is a critical operation in many research laboratories and operations. Much protein analysis is performed using high-pressure liquid chromatography (HPLC) with ultraviolet (UV) or fluorescence detection. Although these methods are robust and widely used, certain issues limit their utility. For example, some proteins or peptides may have a poor UV response and can be difficult to detect. Direct comparisons of protein levels when quantitated directly by UV can also be problematic due to differences in extinction coefficients of various proteins…
The Dale and Betty Bumpers Vaccine Research Center (VRC) at the National Institutes of Health (NIH) was established to facilitate research in vaccine development. The VRC is dedicated to improving global human health through the rigorous pursuit of effective vaccines for human diseases. It was established by former president Bill Clinton as part of an initiative to help develop an AIDS vaccine and is part of the NIAID organization. Since the inception of the VRC, its mission has expanded to include the development of vaccines against bioterrorism and emerging infectious diseases…
R&D output continues to climb with advances in laboratory automation and data analytics but, when measured in terms of the number of drug approvals versus expenditures, the productivity of R&D has been in decline. This paradox confounds many organizations as they struggle to improve. A common strategy has been to invest in informatics. These investments are meant to increase the effectiveness and speed of the decision-making process in which validated findings are compared against predicted outcomes — to close the loop between design and experimentation. But the loop has not been closed in a broad and coherent way, and increased productivity has not been achieved. Instead, the information management landscape in life sciences R&D has remained fragmented, with barriers between disciplines and geographies that prevent broad-based productivity gains…
There is an increasing emphasis in clinical and translational research on the discovery and development of biomarkers that are indicative of a disease state. While biomarkers are not exclusively proteins, the emergence of new mass spectrometry platforms combined with the human genome databases has rejuvenated the search for biomarker proteins, especially in readily available body fluids such as blood. There is currently a tremendous need for an improved ability to “mine” the full depth of the proteome in a high throughput manner. To advance clinical proteomics, methodologies are needed that can accommodate higher throughput while facilitating the ability to observe large numbers of protein events…