Transfection is a technique that makes it possible to modify the genetic content and therefore gene expression of host eukaryotic cells, both in vivo and in vitro. This makes transfection an important tool for drug discovery, the CRISPR-Cas9 technique, studying cell biology, cellular functions and molecular mechanisms of disease. The process centers on inserting proteins, nanoparticles or nucleic acids (such as cDNA, mRNA) into the cytoplasm of cells.
Previously, the use of magnetic beads was limited to small volumes. The difficulty of scaling up beyond a few milliliters was misinterpreted as a limitation of the technology itself. However, as discussed in this e-book, the problem is not the biomagnetic separation process, but a lack of understanding of the physical processes governing it. Once you identify the key parameters that control the magnetic bead's behavior, it is easy to choose the right tools and methods to validate the process and replicate it at different volumes.
Once you have defined the required magnetic force, with a constant magnetic force separation device, it is simple to scale up production. Having validated the magnetic force at a small scale, the same force value can be used for a larger system, even in a different magnetic separation system. Because the conditions remain the same, efficiency (no losses) and batch consistency (no irreversible aggregation) are guaranteed.
When designing a magnetic separation strategy, it is easy to get caught up in the properties of the superparamagnetic beads and how to coat them with the biomolecule of interest (antibodies, antigens, DNA, RNA, oligonucleotides, aptamers...). It is exciting to choose a bead and tailor its surface ligands to perfectly match your target molecule, but don’t stop there! The magnetic separation rack is equally important to a successful identification, isolation, or enrichment protocol. After all, a perfectly designed bead will be useless without a properly designed magnetic rack to efficiently recover it from the solution.
In the recent decades, proteins gained importance after the advent of more advanced analytical procedures and novel genetic or molecular engineering methods. Proteins are cell products and have various physiological functions in the body. Hence, any abnormality in gene expression (mRNA defects), amino acid sequence or structural dysfunction of proteins leads to severe diseases and pathological conditions.
Under constant magnetic force conditions, optical monitoring of the biomagnetic separation process provides information on both when the separation is complete and the characteristics of the magnetic bead suspension.
Protein purification is a fundamental part of studying proteins, peptides, and nucleic acids, necessary for a wide range of clinical, research and industry applications. But choosing the most appropriate protein purification system can be challenging, especially for researchers who are just starting to think about automating their protein purification protocols.
Constant magnetic force separation systems generate the same conditions for all magnetic beads in the suspension. As bead behavior is consistent at every point of the working volume, any changes in the suspension's opacity can be directly related to changes in the suspension’s characteristics.
To ensure a consistent biomagnetic separation process, all the magnetic beads should experience the same conditions. Controlling the magnetic force is key to achieving consistency within and between batches, especially when scaling up. Classical magnetic separators generate a magnetic force that is very high on the side of the vessel closest to the magnet but declines rapidly with distance. The magnetic force experienced by beads in the retention area is therefore greatest, while the beads farthest away experience the lowest force.
Proteins are one of the four macromolecule building blocks of life. The other three are carbohydrates, lipids, and nucleic acids. Proteins are long strings of amino acids that fold together into what are called “hierarchical structures” in order to perform specialized functions within the cells and tissues of all living organisms.