The Progression of the Development of Antibody pY26: Recognizing Phosphorylated Troponin I at the Tyrosine-26 Amino Acid Residue

Biological and Biomedical SciencesAntibodies are used in the western blot technique to identify or quantify a specific protein in a sample. These antibodies cannot be synthesized in a laboratory and must be developed within an animal. It can take months for the antibody to fully mature, and the antibody must be periodically tested to document its progression. The antibody of interest in this study identifies phosphorylated troponin I (TnI) at the Tyrosine-26 (Tyr-26) amino acid residue. The antibody was tested by using four known samples: human phosphorylated TnI, human non-phosphorylated TnI, rat phosphorylated TnI, and rat non-phosphorylated TnI. The antibody was tested at five different time intervals: Pre-bleed, and bleeds 1-4.Theoretically, the antibody would be observed on the phosphorylated samples and would not be observed on the non-phosphorylated samples. As time progresses, the antibody should also become more specific (binding only to phosphorylated TnI), and more intense (binding to phosphorylated TnI more frequently). As each subsequent test bleed was used on the samples, the specificity and intensity of the antibody increased. Further work was done on the antibody in terms of determining what concentration it should be used at to create the most clear images. When used at a 1:60,000 antibody:TBS + 0.1% Tween ratio, the image of the blot is the most clear. With an effective antibody that identifies phosphorylated TnI Tyr-26, unknown samples can be tested for the phosphorylation. Trends between heart failure and phosphorylation of the residue can be established and ultimately treatments can be created depending on the effect of the phosphorylation.Undergraduate Research OfficeAcademic Major: Biolog