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.
Mass spectrometry is a technology which is widely used in most scientific disciplines that require accurate and precise measurement of elemental and molecular components. Its use in the pharmaceutical sector is often associated with the Drug Discovery and Development process. The primordial step to analyze a sample is to perform a sample treatment to enable its study. It really requires know-how to obtain good quality results. The following tips and tricks can help you to rapidly select a protein purification strategy and to anticipate some possible problems.
Decidedly, producing and purifying a protein is a form of art.
Indeed, each recombinant protein (rec. protein) to be produced in vitro has specific characteristics and singularities that make its production and purification most definitely challenging. In this post, I’d like to share and review a few tips and basics for optimal design of experiment when producing functional recombinant proteins and working on protein purification.
Protein biochemistry brings together a vast and varied world of methods of protein production, purification and characterization. Once you have successfully achieved the production of your protein in a selected system, you need to think about the following steps. Indeed, the quest does not stop there! The next step is purification, during which you will try to isolate your protein of interest from the surrounding contaminants while keeping it soluble and active. Your ultimate aim is to keep your protein “happy” by choosing the perfect buffer. You will be faced with a bewildering array of choices leading you to ask yourself “where do I start…?”.
Keep calm – it’s easy… [Read more…]
Gel filtration (GF), also referred as Size Exclusion Chromatography (SEC), plays a key role in the high quality purification of enzymes, polysaccharides, nucleic acids, proteins and other biological macromolecules. Gel filtration is the simplest and mildest of all chromatography techniques to separate biomolecules on the basis of difference in size. Nevertheless, the list of available columns is quite awesome and the characteristics of each of them are very different. Also, each characteristic highly influences the quality of the final purified product with important consequences in downstream applications.
I’ve put together a simple guide to help you find your GF system, together with some tips and tricks based on my experience when producing recombinant proteins and antibodies for our clients involved in the early R&D stages through to the latest phases of the bio-production flow.
Download your copy of the guide “Gel Filtration – which column should you choose for your size exclusion purification?“
If you have any questions or if you need some help, leave a message below to get in touch with our lab experts!
…or how to make your protein attractive
Biomagnetic separation is a powerful lab tool in protein biochemistry which can be used in a wide range of applications including analysis tools (for protein solubility and protein-protein interaction studies, and for testing several purification buffers…), high throughput screening (HTS) and larger protein purification scale.
Indeed, it’s an easy, quick and efficient method to purify recombinant proteins and antibodies. Note that it also can be used to remove endotoxins and abundant protein contaminants from samples. Let’s take a look together at the large number of possibilities available and how we can help you to design the most adapted protocol for your study.
Monoclonal antibody (mAb) engineering and their development as therapeutic or diagnostic tools have become the must in pharmaceutical industry, in medical biotech and in the research world. Purification of this type of biosimilar requires a particular know-how.
Because of their rapid growth and their affordability, bacterial expression systems are very convenient when producing recombinant proteins.
Nevertheless, one of the key elements in the design of the experimental procedures for producing a recombinant protein in E. Coli is the selection of the right bacterial strain for the right protein. This is not an easy step but remains of utmost importance.
Based on our in-house experience, we’ve compiled a guide aimed at helping you in your choice of the “ideal” engineered bacterial strain for efficient recombinant proteins expression.
You’ll find useful information for selecting the reliable strains suited to your downstream applications, with characteristics (quality, biological activity, folding…).
Interested? Download your copy here:
Guide to choosing the best bacterial strains for recombinant protein expression
Pharmaceutical, biotech and medical device industries apply restrictive quality systems in their bio-production processes. Among the numerous parameters controlled daily by scientists, I have selected a particular one for this post, i.e endotoxin detection.
Isolation of highly purified untagged proteins is crucial in today’s R&D programs. Up to now, recombinant protein production approaches were often based on the use of “tags” to make protein purification (and solubility) more convenient. Unfortunately, these tags can be real experimental hurdles in downstream applications. Protein experts are now considering “tag-free” alternatives (including outsourcing) for producing untagged proteins more rapidly, more efficiently, and without jeopardizing the rest of the research project. Let’s see how this is possible.