A Trypan Blue-Based ELISA Practice Test to Evaluate Trainee Performance

Abstract

Enzyme-linked immunosorbent assay (ELISA) is a valuable tool in various chemical and biological fields for investigating the presence and quantity of proteins of interest in given samples. The high importance of the assay makes it widely used in biological research and hence a crucial skill to master for up-and-coming scientists. A major impediment to teaching the skill to new students in research laboratories is the high cost of commercial ELISA kits. As a result of the cost of the kits, most investigators prefer that more experienced students or technicians carry out the assay to prevent errors, thus depriving less experienced students the chance to actively learn the process. Here, we present a practice ELISA test that can be done in any laboratory using different dilutions of trypan blue as the reagents and samples, thereby creating a product at one-hundredth the cost of the least expensive ELISA kits. Research has continuously shown that active learning tools such as this simulation enhance student engagement and self-efficacy. This practice test also generates a numerical score based on the students’ accuracy in pipetting and attention to detail, allowing students to identify sources of error. Therefore, the practice ELISA experiment acts as a medium to teach proper technique before performing a real ELISA, thus avoiding wasting expensive reagents and limited samples.