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Published on: January 6, 2016
Application of sandwich ELISA for detecting tag fusion proteins in high throughput
Zhu-wei Xu1, Tao Zhang, Chao-jun Song
1Department of Immunology, The Fourth Military Medical University, 17 ChangleWest Road, Xi'an, 710032, Shannxi, China.
Based on a series of mAbs against four frequently used tags--the human Ig Fc fragment, GST, maltose-binding protein, and thioredoxin--we developed corresponding sandwich enzyme-linked immunosorbent assay (ELISA) to detect these tag fusion proteins. As a supplement for Western blot, the successfully established ELISA was specific, sensitive, quantitative, easy to perform, time-saving, and last but not least, suitable for high-throughput screening of tag fusion proteins. Determination of soluble tag fusion proteins expressed by various systems with the sandwich ELISA developed in the present study could be a valuable and promising tool for the wide application of tag-protein fusion systems in the rapidly growing field of proteomics research.
Based on a series of mAbs against four frequently used tags--the human Ig Fc fragment, GST, maltose-binding protein, and thioredoxin--we developed corresponding sandwich enzyme-linked immunosorbent assay (ELISA) to detect these tag fusion proteins. As a supplement for Western blot, the successfully established ELISA was specific, sensitive, quantitative, easy to perform, time-saving, and last but not least, suitable for high-throughput screening of tag fusion proteins. Determination of soluble tag fusion proteins expressed by various systems with the sandwich ELISA developed in the present study could be a valuable and promising tool for the wide application of tag-protein fusion systems in the rapidly growing field of proteomics research.
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Tagging and Fusion Proteins
Enzyme-Linked Immunosorbent Assay
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

