Introduction: (A) Elisa, which stands for Enzyme-Linked Immunosorbent Assay, is a test that can detect diseases in blood and bodily fluids using antigens that bind to the foreign antigens. When antigens trigger an immune system response, millions of antibodies find the antigens and bind to it. The secondary antigens that are enzyme linked are the next ones to bind to the antigens. Next, Chromogenic enzyme substrate is added which will color the wells. If they remain colorless, then they are negative, and if colored, the results are positive. Today, ELISA is used for many different purposes which include pregnancy tests, detecting drug use and disease in people, reassuring that food is labeled correctly and testing indoor air quality. With its rapid test results, ELISA has been very helpful for many diverse purposes. A very important aspect of the ELISA test is the antibodies. They prove to be useful because they can bind really well (with antigens). Some different uses of binding are immunostaining, immunoblotting or western blotting. Different kinds of cells (such as cancer cells) may be identified by the process of immunostaining. A protein's size and quantity may be determined by immunoblotting or western blotting. Antibodies can also be manufactured. Polyclonal antibodies are made by immunizing an animal. An animal (most likely a goat, rabbit, or sheep) would be injected with a purified protein that would allow the animal to make different antibodies to develop within the animal. Serum can then be collected by drawing blood from the animal and removing the blood cells from it. The serum that is isolated from the blood sample of the animal is called antiserum. The final step is to separate the antibodies from the antiserum. These antibodies are called polyclonal. The only down side to polyclonal antibodies is that no two batches will ever be the same. (B) The purpose of the experiment is to find out which two people in the class were originally diseased. This will be done by having everyone share fluids with three other people. After the ELISA test, the results will help to determine who the two disease spreaders were. In reality, the sharing of bodily fluids can be quite common. Someone may cough or sneeze on you with out you knowing and before you know it, you have become a victim to their cold they have shared with you. One in four teenage girls have an STD (from Mr. Chugh's stat of the day) and ELISA would be a way for them to find out. (C) On the first day of the lab, we will obtain a tube of some "bodily fluids". Then, with three other people, we will share our bodily fluids we got and record their tube number and the time at which the sharing occurred. On the second day, we will receive twelve wells which we will add positive/negative controls, and our shared fluids to. It will then be cleaned out with wash buffer. The next step will be to add the primary antibodies then it will be cleaned out with wash buffer. The secondary antibodies are added next and then also cleaned out again. After it is rinsed with the wash buffer, substrate will be added and show the results. (D) I'm not really able to predict if i will be infected or not because the two diseased people could be anybody. It depends on who I share my fluids with. There are two controls in this experiment: a positive and a negative. The positive wells will be added with fluids that are diseased and should show up as blue. The negative wells will be added with fluids that are diseased and should be clear. For the results in our wells to be sufficient, the positive and negative controls must come out as expected when the substrate is added to it.
Results/Observations: After adding substrate, all twelve of the wells appeared to remain the same. This might mean a source of error occurred while the lab took place. Therefore, it remains a mystery if my lab group and I have been infected.
Discussion: With missing data from table four and five, it is almost impossible to determine who is the original diseased people are. The originally diseased people did share very quickly though because everyone is infected except for a few. The source of error in our lab was washing out the wells before adding the positive/negative controls and our fluids. The primary antigens did not have a chance to bind which made it impossible for the secondary antibody to bind and so on.
