Jun, 2012
Epitope analysis using membrane-immobilized avidin and protein A
PROTEIN EXPRESSION AND PURIFICATION
- ,
- ,
- ,
- Volume
- 83
- Number
- 2
- First page
- 177
- Last page
- 181
- Language
- English
- Publishing type
- Research paper (scientific journal)
- DOI
- 10.1016/j.pep.2012.03.017
- Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
Adrenocorticotropic hormone (ACTH) and transferrin were trapped by biotinylated anti-ACTH antibody and anti-transferrin antibody, respectively, bound to membrane-immobilized avidin. Polypeptides with the sequences SYSMEHFR. SYSMEHFRWGKPVGK and SYSMEHFRWGKPVGKK were bound to the biotinylated anti-ACTH antibody on the membrane-immobilized avidin after the trapped ACTH was digested with trypsin on the membrane and non-binding polypeptides were washed from the membrane. Further, the polypeptides with the sequence SYSMEHFRWGKPVGK and SYSMEHFRWGKPVGKK were trapped by anti-ACTH antibody bound to membrane-immobilized protein A. The antibody recognized the WGKPVGK region of the antigen, ACTH. Polypeptide with the sequence SMGGKEDLIWELLNQAQEHFGKDK was bound to the biotinylated anti-transferrin antibody on the membrane-immobilized avidin after the trapped transferrin was digested with trypsin on the membrane and non-binding polypeptides were washed from the membrane. Further, the polypeptide with the sequence SMGGKEDLIWELLNQAQEHFGKDK was trapped by anti-transferrin antibody bound to membrane-immobilized protein A. The antibody recognized the SMGGKEDLIWELLNQAQEHFGKDK region of the antigen, transferrin. These results thus indicate that the combined methods of membrane-immobilized avidin and protein A can be applied to examine the epitopes of antigens. (C) 2012 Elsevier Inc. All rights reserved.
- Link information
- ID information
-
- DOI : 10.1016/j.pep.2012.03.017
- ISSN : 1046-5928
- Web of Science ID : WOS:000305166300010