Enzyme-linked immunosorbent assay (ELISA) is a widely used diagnostic tool in basic and applied research. This project is to develop the reagents and protocols for an ELISA for a biochemistry undergraduate laboratory class. Currently, biochemistry lab students study protein purification by purifying a form of green fluorescent protein (GFP), the expression product of pGLO plasmid from Bio Rad Laboratories. The next desired lesson would be to learn about ELISAs in various forms of this technique, including direct, indirect, and sandwich, and competitive formats. Detection of GFP will be the model analyte for these studies. Knowledge and practice with ELISA formats are important skills for biochemistry students to learn for future academic and industrial research. For the development of this protocol, it is necessary to express and purify significant quantities of green fluorescent protein (GFP), as well as prepare reagents for the ELISA protocol such as, enzyme-linked conjugates of GFP, anti-GFP rabbit polyclonal antibodies, and goat anti-rabbit enzyme linked polyclonal antibodies. It is essential to optimize concentrations, wash steps, and incubation times for ELISA formats. The final step will be to write the protocols that will be used for biochemistry students in the lab class.

Designing an ELISA for biochemistry laboratory students to study immunochemistry in biotechnology

251st National Meeting (2016)

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