Application of sandwich ELISA for detecting tag fusion proteins in high throughput.
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.
Xu
Zhu-wei
ZW
Department of Immunology, The Fourth Military Medical University, 17 ChangleWest Road, Xi'an, 710032, Shannxi, China.
Zhang
Tao
T
Song
Chao-jun
CJ
Li
Qi
Q
Zhuang
Ran
R
Yang
Kun
K
Yang
An-gang
AG
Jin
Bo-quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins