Enzyme-Linked Immunosorbent Assay (ELISA) has continued to grow in use for carrying out assays globally. The enzyme-linked immunosorbent assay (ELISA) is used to test for the presence of a specific protein or polypeptide in biological fluids, such as blood serum, urine, and cerebrospinal fluid. For example, ELISA can determine if antibodies are present that are specific for a certain antigen.
The ELISA assay uses a microtiter plate that has been coated with an antigen of interest. The plate will have many microscopic wells that contain the antibody of interest. A sample containing the protein or polypeptide of interest is added to each well and allowed to react with the antibodies on the plate. If there is an antigen in the sample, it will bind to the antibody attached to the well plate creating an immune complex. After washing away unbound materials, an enzyme-linked secondary antibody will be added. The enzyme conjugated to this antibody will react with a substrate that produces a colorimetric or fluorescent signal when acted upon by this enzyme. This allows for easier detection of the immune complex by observing the colored or fluorescent reaction products.
An enzyme-linked immunosorbent assay (ELISA) using glycoprotein was developed to determine human seminal plasma vitellogenin (VTG). The ELISA was performed in a microtiter plate, and the absorbance was measured with a microplate reader. It has been previously established that the enzyme-linked immunosorbent assay (ELISA) using glycoprotein is a sensitive and specific method for determining the presence of antibodies to the herpes simplex virus, type 1 (HSV-1).
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What is Glycoprotein?
Glycoprotein is a protein that has sugar chains attached to it. Glycoproteins are sugar-protein combinations and are bound with sugars or saccharides. The glycoprotein combination is important in the functioning of human, animal, and plant cells. Glycoproteins are involved in many types of functions in our body. Some of which include helping the blood clot, without which we would bleed to death, and regulating some hormones. Glycoproteins are also found on the surfaces of cells where they protect against infection by bacteria and viruses, and they are used by immune cells to detect foreign invaders. Glycoproteins even have an important role in fertilization and are present during pregnancy.
Glycoproteins serve as growth factors, stimulating cell division in various body organs. Glycoproteins act as receptors for specific molecules that recognize harmful microorganisms. Some glycoproteins, such as those on the cell membrane, play a role in preventing harmful microorganisms from entering the body or spreading throughout it.
There are three major glycoprotein types: serine protease inhibitors, lectins/carbohydrate-binding proteins, and sialic acid-binding lectins. These proteins are found on the surface of most animal cells and function as part of the immune system to shield the host from pathogens.
Glycoprotein Based ELISA Assay Analysis
Glycoprotein analysis has become a key tool for biomedical research because of its sensitivity and specificity in identifying particular proteins and determining protein structure. The use of glycoproteins in ELISA is based on the ability of these proteins to bind carbohydrates. Glycoproteins interact specifically with their ligands, a carbohydrate or other type of molecule that contains a carbohydrate or noncarbohydrate component.
In glycoprotein ELISA testing, the PSA protein is often used as the “glycoprotein.” In this case, the PSA protein will be attached to a piece of glass. The glass slides are coated with antibodies that can detect the presence of PSA protein in the blood. These antibodies will attach themselves to the PSA protein when it comes into contact with them, creating a reaction between them. The results of glycoprotein ELISA testing are often compared to standard ELISA tests.
Several ELISA kits work with glycoprotein; we would be looking into some of them: MAG ELISA, EPO ELISA, and laminin antibody.
Human MAG ELISA
Human MAG Elisa has 96 wells per kit with removable strips. Human MAG Elisa has reactive species as humans a d has a reactivity of 50pg/ml. It has an assay range of 312 pg/ml – 20,000 pg/ml and has no detectable cross-reactivity with other relevant proteins. Human MAG Elisa should be Stored at 4°C for 6 months and at -20°C for 12 months. Avoid multiple freeze-thaw cycles.
Human MAG PicoKine ELISA Kit Standard Curve (https://www.bosterbio.com/human-mag-picokine-elisa-kit-ek2061-boster.html)
EPO ELISA
Epo Elisa, also known as Human Erythropoietin, has 96 wells per kit and removable strips. Epo Elisa has its reactive species as humans and has a sensitivity of 1mIU/ml an assay range of 3.12 mIU/ml – 200 mIU/ml. The same types for EPO Elisa include cell culture supernatants, serum, and plasma (heparin, EDTA).
Human EPO PicoKine ELISA Kit Standard Curve (https://www.bosterbio.com/human-epo-picokine-elisa-kit-ek0332-boster.html)
Anti-Laminin Antibody
This has a size of 100μg/vial with the mouse as its host and humans and Pigs as the reactive species. The antibody is monoclonal in clonality and has Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as a preservative. Anti Laminin antibody should be stored at -20˚C for one year from the date of receipt. After its reconstitution, it should be stored at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Finally, avoid repeated freeze-thaw cycles.
Booster kit box (https://www.bosterbio.com/anti-laminin-antibody-monoclonal-ma1054-boster.html)