Essentially, MyBioSource, Inc. is a biotechnological product distribution company formed to make a large portfolio of laboratory research reagents, both hard to find and common items, with wide-reaching distributions.

Backed by an elite network of laboratories and manufacturers, the company was launched in 2007 in Vancouver, British Columbia, Canada. Their headquarters and operations were repositioned to San Diego, United States

Amazingly, the company distributes a number of science reagents and assay kits which include custom recombinant proteins and antibodies, real-time PCR and quantitative ELISA kits. The current catalog of the company lists over 1 million individual products used in biotechnological,   pharmaceutical industries.

And at this moment, we’ll get to know more about MyBioSource’s ELISA Kits.

Defining Elisa Kits

Little did everyone know, the enzyme-linked immunosorbent assay (ELISA) is an ordinarily used analytical biochemistry assay. The assay uses a solid-phase enzyme immunoassay (EIA) to detect the presence of a ligand (usually a protein) in a liquid sample using antibodies directed counter to the protein to be measured.

The ELISA kits have been used as a diagnostic tool in medicine, plant pathology, and biotechnology, as well as quality control,  check in various industries.

In the simplest form of an ELISA, antigens from the sample are attached to a surface. Then, a matching antibody is applied over the surface so it can bind to the antigen. This antibody is associated with an enzyme, and in the final step, a substance comprising the enzyme’s substrate is added. The subsequent reaction produces a detectable signal, most commonly a color change in the substrate.

Moreover, performing an ELISA involves at least one antibody with specificity for a particular antigen. The sample with an unknown amount of antigen is immobilized on solid support usually a polystyrene microtiter plate either non-specifically via adsorption to the surface.

After the antigen is immobilized, the detection antibody is added, forming a complex with the antigen. The detection antibody can be covalently linked to an enzyme or can itself be detected by a secondary antibody that is linked to an enzyme through bioconjugation.

Between each step, the plate is typically washed with a mild detergent solution to remove any proteins or antibodies that are non-specifically bound.  Furthermore, after the final wash step, the plate is developed by adding an enzymatic substrate to produce a visible signal, which indicates the quantity of antigen in the sample.

Worth mentioning, ELISA can perform other forms of ligand- binding assays instead of strictly “immuno” assays, though the name carried the original “immuno” because of the common use and history of the development of this method.

Fundamentally, the technique essentially requires any ligating reagent that can be immobilized on the solid phase along with a detection reagent that will bind specifically and use an enzyme to generate a signal that can be properly quantified.

In between the washes, only the ligand, and its specific binding counterparts remain specifically bound or “immunosorbed” by antigen-antibody interactions to the solid phase, while the nonspecific or unbound components are washed away.
Unlike other spectrophotometric wet lab assay formats where the same reaction well can be reused after washing, the ELISA plates have the reaction products immunosorbed on the solid phase, which is part of the plate, and so are not easily reusable.

Use of Elisa Kits

Since the ELISA can be executed to evaluate either the presence of antigen or the presence of antibody in a sample, it is a useful tool for determining serum antibody concentrations such as with the HIV test or West Nile virus.

It has also found applications in the food industry in detecting potential food allergens, such as milk, peanuts, walnuts, almonds, and eggs and as a serological blood test for coeliac disease. ELISA can also be used in toxicology as a rapid presumptive screen for certain classes of drugs.
In addition, the ELISA was the first screening test widely used for HIV because of its high sensitivity. In an ELISA, a person’s serum is diluted 400 times and applied to a plate to which HIV antigens are attached. If antibodies to HIV are present in the serum, they may bind to these HIV antigens. The plate is then washed to remove all other components of the serum.

A specially prepared “secondary antibody” which is an antibody that binds to other antibodies is then applied to the plate, followed by another wash. In connection, this secondary antibody is chemically linked in advance to an enzyme.

Consequently, the plate will contain the enzyme in proportion to the amount of secondary antibody bound to the plate. A substrate for the enzyme is applied, and catalysis by the enzyme indicates a change in color or fluorescence. ELISA results are reported as a number; the most controversial aspect of this test is determining the “cut-off” point between a positive and a negative result.

It’s Standards

As an analytical biochemistry assay, ELISA involves detection of an “analyte” in a liquid sample by a method that continues to use liquid reagents during the “analysis”  that stays liquid and remains inside a reaction chamber or well needed to keep the reactants contained and  it is opposed to “dry lab” that can use dry strips – and even if the sample is liquid, the final detection step in “dry” analysis involves reading of a dried strip by methods such as reflectometry and does not need a reaction containment chamber to prevent spillover or mixing between samples.

As a heterogeneous assay, ELISA splits some component of the analytical reaction mixture by adsorbing certain components onto a solid phase which is physically immobilized.

 In ELISA, a liquid sample is added onto a stationary solid phase with special binding properties and is followed by multiple liquid reagents that are sequentially added, incubated and washed followed by some optical change (e.g. color development by the product of an enzymatic reaction) in the final liquid in the well from which the quantity of the analyte is measured.

The quantitative “reading” usually based on detection of the intensity of transmitted light by spectrophotometry, which involves quantitation of transmission of some specific wavelength of light through the liquid (as well as the transparent bottom of the well in the multiple-well plate format). The sensitivity of detection depends on the amplification of the signal during the analytic reactions.

In the meantime, enzyme reactions are very well known amplification processes, the signal is produced by enzymes which are linked to the detection reagents in fixed proportions to allow accurate quantification – thus the name “enzyme-linked”.
Conventionally, like other forms of immunoassays, the specificity of antigen-antibody type reaction is used because it is easy to raise an antibody specifically against an antigen in bulk as a reagent. Otherwise, if the analyte itself is an antibody, its target antigen can be used as the binding reagent.

Takeaway

To sum up, MyBioSource is really beneficial. With its products, these lead everyone to be better. So what are you waiting for, avail now!