Microbeads are composites of nanosized magnetic particles embedded in a non-magnetic matrix, the size of each bead in the range of micrometer diameter. Microbeads were originally the discovery of John Ugelstad, who was able to create uniform polystyrene spheres. Magnetic microbeads are superparamagnetic microbeads that specifically only become magnetized in the presence of a magnetic field, and this is a reversible phenomenon. These microbeads, made of materials with magnetic properties such as magnetite, have several uses in scientific research. An important use of magnetic microbeads is for separation of biological molecules from mixtures.
Magnetic bead technology can be used to separate several types of biological molecules from the solution which they are in. You can separate a protein of interest or a nucleic acid, DNA or RNA, from a heterogeneous mixture such as cell lysate. There are kits and protocols that have been established and are commercially available for isolating DNA or RNA from a solution. These kits contain buffers that can make the isolation process time efficient, and keep the yield of nucleic acid recovery high. Beyond that, some laboratories are investigating an optimization for isolating both DNA and RNA with a protocol that is even more time and cost effective.
Enzymes play a surprisingly important role in modern industry, and are essential to the production of more commercial products than one would initially consider. Enzymes are proteins that speed up reactions and improve yield by increasing available precursors for downstream reactions. Perhaps the most obvious use for enzymes in industryisthe production of cheese, bread, and alcohol. In these traditional applications the enzymes are part of microbial machinery such as bacteria or yeast. Over time scientists have been able to isolate specific enzymes and to understand their catalytic functions well enough to incorporate them with or without their microbial hosts into a wide variety of somewhat surprising situations. For example, enzymes are used in the production of textiles, detergents, biofuels, and pharmaceutical products. Large quantities of desired enzymes are required for these applications, and they need to be available in the purest form possible. The purity of enzymes in industry is particular important for pharmaceutical applications where the products as well as the process are susceptible to review and control by regulatory associations. Batches of enzymes in industry undergo regular process validation to ensure batch-to-batch consistency.
The industrial centrifuge plays an integral role in the production of more things than one would initially expect. It is a commonly used tool in the food and agricultural sector, At pharmaceutical and biotechnology companies, for environmental management, and in the chemical industry. The word industry conjures up images of combination and creation—adding materials together to produce a final product. However, the separation of materials is just as important as the combination of materials. We can't create a new product until we have pure reactants to work with. This is especially important in the pharmaceutical and biotechnological realms, where reactant purity is essential to the production of a product that is safe for human consumption. This is where the centrifuge comes in. The centrifuge is used to separate heterogeneous mixtures into components varying by density.
An elution buffer plays an essential role in every immunoprecipitation protocol or assay that requires the release of a target antigen from a capture antibody. Elution buffers are necessary in protocols utilizing a stationary affinity column, and are also required in protocols using mobile solid supports in solution.
A cell lysis buffer is a critical first component to any isolation protocol. It is fundamental to the first step of protein or nucleic acid extraction as it aids in the chemical breakdown of cell membranes and compartments, enabling target molecules to escape. There are many types of lysis buffers; most are easy to make, but most are also commercially available. They are often included in kits for immunoprecipitation, co-ip protocol, nucleic acid isolation, and others. When using a lysis buffer for protein capture it is a good idea to add protease inhibitors prior to use in order to protect proteins.
The sandwich ELISA is a type of Enzyme-linked immunosorbent Assay that uses two antibodies: a capture antibody and a detection antibody. The purpose of any ELISA is to detect the presence of a target antigen in a sample. In a sandwich ELISA the target antigen is bound between a capture antibody and a detection antibody. The capture antibody is immobilized on a surface, while the detection antibody (conjugated to an enzyme or fluorophore label) is applied as a last step before quantitation.
The force of gravity will cause sedimentation of particles from a heterogeneous mixture; larger and denser particles sedimentfaster than the smaller and less dense particles. This phenomenon is useful for separating heterogeneous solutions into independent components, and for the isolation and enrichment of target molecules, cells, and cell organelles. Differential centrifugation accelerates the separation process by introducing centripetal forces many times greater than gravity. The precipitated particles form a pellet at the bottom of the tube during centrifugation. The rate of sedimentation is dependent on the size and density of the particles, so centrifugation can be used to isolate target particles simply by controlling centrifugal force or the rate of centrifugation. The rate of centrifugation is reported as angular velocity by revolutions per minute (rpm) or as acceleration(g). RPM is dependent on the radius of the rotor in the centrifuge.
In the fight against COVID-19, testing of patient samples has been mostly conducted using standard techniques, which has kept clinics struggling to keep up with the demand for testing. The first step in coronavirus testing that needs to be more efficient is the RNA extraction.
Introduction to Chemiluminescence immunoassays
Serological tests are used to gain a deeper understanding of the immune response to pathogens and the tests help maintain community health by checking for antibodies in human biological samples. Chemiluminescence is a widely used system of reporting binding events. It is preferred because it uses a simple device for measurement, often one that measures output of visible light. This also allows the process to have a wide dynamic range, detecting light from binding events whether the sample is dilute or concentrated. Such detection is done with high sensitivity and with low background noise. The chemiluminescent magnetic microparticle immunoassay (CMIA) is a method developed to bring together the advantages of chemiluminescence and magnetic particles for immunoassays.