Examining Inhibition of Elastase via a Radial Diffusion Assay

Abstract

Alpha-1-antitrypsin deficiency (A1AD) is a disorder inherited through certain genotypes and is in about 1 in 2500 individuals . This disorder decreases levels of alpha-1 in the bloodstream which, in turn, leads to an unchecked amount of neutrophil elastase. The purpose of elastase, as is the case with most enzymes, is to break down its corresponding protein in this case, elastin, which is found in great quantities in the body’s organs, namely, the lungs. People who are born with A1AD are usually susceptible to lung diseases such as emphysema and COPD and in rare cases, liver disease. Rather than trying to cure the disorder itself, what we hope to accomplish in this research is to find an inhibitor that would appreciably prevent the spread of elastase throughout tissue containing elastin. To determine this, we will be looking at the spread of elastase through individual wells filled with elastin. Each well on the plate will have a small depression hollowed out in the middle and be filled with different amounts of elastase and inhibitor, the different amounts of elastase representing the different ways that A1AD can affect different genotypes. Each well will be watched for different amounts of time. The effectiveness of the inhibitor will be measured by the radius out from the depression site; the smaller the radius, the more effective the inhibitor. We are hoping that this study will lead into treatment for this symptom of the disorder.