A simple, economical and effective ELISA using simulated serum sample for teaching the concept and application of antigen–antibody interactions in undergraduate laboratory

ABSTRACT

Enzyme-Linked Immunosorbent Assay (ELISA) is a versatile and widely used technique to detect and quantify small molecules in various clinical and analytical applications. It is also an effective tool for demonstrating the specificity between antibody and antigen to students across diverse disciplines. However, undergraduate laboratories often face the difficulty of performing proper ELISA exercise due to time and facility constraints. Here, we report a cost-effective, technically simple ELISA exercise which can be completed within 2.5-h at room temperature. This microtitre plate-based ELISA exercise utilises easily sourced inexpensive materials and simulated serum samples which are safe to handle in the undergraduate lab settings. The hands-on procedure and a real-life scenario of disease (Rheumatoid Arthritis) diagnosis reinforce students’ understanding of antigen–antibody interactions. ELISA results are displayed by a colourimetric enzymatic reaction, which can be analysed by eye without specialised equipment. This laboratory exercise was designed as part of the final year’s immunology course in a three-year undergraduate Biomedical Science programme, but can also be adapted to suit a broad range of student skill levels in science-fields because of its easy set-up and low technical requirement. The procedure and scenario can be also adjusted to meet the specific needs of other educator’s goals.

KEYWORDS:

• Antigen-antibody interactions
• hands-on ELISA exercise
• detecting specific antibody
• enzymatic immunologic reaction

Acknowledgments

We would like to thank Associate Professor Roger Booth, at the University of Auckland, New Zealand for designing the original student-protocol of this exercise, Emeritus Professor Phil Crosier, at the University of Auckland, New Zealand for the encouragement to develop the protocol to prepare simulated serum samples, and Dr Chris Hall, Senior Lecturer at the University of Auckland, New Zealand for his critique and comments on this manuscript.

The Authors have no conflicts of interest to declare.

Disclosure statement

No potential conflict of interest was reported by the authors.

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Additional information

Notes on contributors

Latifa B. Khan

Latifa B. Khan, Senior Teaching Technician Created, tested, and optimised the preparation protocol of ELISA exercise. Optimised the student’s laboratory protocol, and organised the teaching of the laboratory exercise. Planned the work that led to the manuscript production. Wrote the initial draft, reviewed successive versions and approved the final version of the manuscript.

Jia-Yun C. Tsai

Jia-Yun C. Tsai, Research Fellow Participated in teaching and demonstrating the laboratory exercise. Reviewed successive versions, and approved the final version of the manuscript.