Recombinant protein expression and purification for therapeutic development or for in vitro studies can sometimes be a real challenge. It requires huge investments in both time and cost in order to obtain high yields of pure and active recombinant proteins. My aim in writing this post, is to give you a simple guide to easily establish optimal conditions for recombinant protein expression and production in E. Coli, of problematic unstable proteins of interest.
Kupffer primary cells are macrophages endogenous to the liver which have the ability to modulate hepatic inflammation and injury associated with various pathophysiologies and toxicities. Pro-inflammatory cytokines released by activated Kupffer cells, such as TNF-α and IL-6, are associated with up-regulation of acute-phase response proteins and suppression of CYP enzymes. For new biological entities, particularly immunomodulators, evaluating the potential for Kupffer cell activation is an emerging concept in preclinical development. Kupffer cells are estimated to comprise approximately 4-8% of total liver cell content and approximately 20% of non-parenchymal cells (Raccanelli and Rehermann, 2006). [Read more…]
As is already known, multiple families of small GTPases (Ras, Arf and Rho families) make up the Ras GTPase superfamily. Due to their role as molecular switches in cell signalling (active GTP-bound state vs inactive GDP-bound states) and in many other cellular responses (cytoskeletal reorganization, regulation of transcription, apoptosis…), these small GTPases are the subject of intense investigation to try to understand the mechanisms that regulate activation and inactivation of this proteins.
In this post, I invite you to discover a revolutionary way to study these cell signalling events by quantifying the level of active small GTPases in your experimental model.
In the field of neurosciences, increasing research is being undertaken on neurodegenerative disorders like Parkinson’s and Alzheimer’s diseases, as well as on nerve injury and regeneration. One of the challenges researchers face is finding the appropriate cellular neuromodels, as close as possible to the in vivo environment. Primary cells are one of the answers, increasingly used in co-culture systems and 3D models. However, cells from the nervous system are not readily accessible, nor are they easy to culture.
Cell Applications Inc. provide solutions to face these challenges, ranging from highly pure, low passage ready-to-use primary cells, to stem cells and induced pluripotent stem cells (iPSC) derived from a variety of clinically relevant species. Let’s take a look at a selection of their neuron, microglia and astrocyte models in relation to their potential applications.
Autologous tumour cell vaccines vs. dendritic cell vaccines: What’s best, and why?
Understanding the best approach for tumour vaccinations is of growing importance in the personalized cancer immunotherapy field. Current models use a variety of treatments, but two include the use of irradiated autologous tumour cells (TCV), or autologous dendritic cells loaded with tumour associated antigens (DCV). In a randomized trial of metastatic melanoma patients undergoing either TCV or DCV, DCV patient survival was far superior at 43.4 vs 20.5 months. Why was the response more effective when the same antigens were provided to the immune response? Dillman, Robert O., et al. brought some answers in a paper published in the Journal for immunotherapy of cancer in 2018). Looking at the cytokine “milieu” with the Quantibody 200 Biomarker Microarray Service helped the authors in understanding the differences in vaccine responses. [Read more…]
For many pathologies such as cancer or neurological disorders, it has been found that Histone Deacetylase (HDAC) activity is disrupted, making them promising targets for drug development. Scientists need access to a broade range of products ( Active Enzyme, inhibitors/activators, Biochemical screening assays…), to simplify their research and identification of HDAC inhibitors.
Rockland Immunochemicals, Inc., (Rockland) provide the highest quality antibodies to the academic, biopharma, and diagnostic industries for use in basic research, assay development, preclinical and clinical studies, and bioprocessing. With facilities in Pennsylvania established since 55 years ago, Rockland manufactures products ideally suited for integration into critical assays such as western blotting, immunohistochemistry (IHC), immunofluorescence microscopy (IF), ELISA, flow cytometry, and 2D imaging. Their antibodies show remarkable quality and reproducibility from batch to batch. Their popular anti-RFP is one example out of many.
Demonstrating Rockland’s commitment in this matter (manufacturing antibodies of high quality and reproducibility), Rockland’s CSO, Dr. Carl Ascoli, testified at the US National Academies of Sciences, Engineering and Medicine’s third of six public sessions on Reproducibility and Replicability in Science on April 18th this year. [Read more…]
I am following up on my series of posts based on the work done by Chris Bohl and collaborators at Sekisui Xenotech. Here, he looks at the effect of cryopreservation on pooled hepatocytes metabolic activities, and how the patented Cryostax technology can address this issue.
Membrane transporters can have clinically relevant effects on the pharmacokinetics and pharmacodynamics of a drug in various organs and tissues by controlling its absorption, distribution and elimination. Together with metabolizing enzymes, they can drive a drug’s pharmacological action, as well as a drug can modulate transporter expression or activity, hence the importance of evaluating transporter-mediated drug-drug interactions recommended by the FDA guidelines. Genomembrane have developed transiently expressing ready-to-use cells to study FDA approved transporters (TransiPort). I had described this product range and their unique features in a previous post. I would like to introduce in this one 2 newly released cells overexpressing OATP1A2 and OATP2B1 transporters. [Read more…]