Protein analysis
Epitope Mapping
Innovative Technology Support Group for Life Science and Drug Discovery
EP-101 : Epitope Mapping
Test Introduction
Epitope is an antigenic determinant which is a structure recognized by an antibody. To confirm the binding position of the antigen and the antibody three analytical methods are performed. Generally, analysis methods are consist of Covalent labeling, Cross Link, Immuno-affinity peptide screening. Three types of analyses are used to determine the linear and conformational epitope positions of the antigen protein.
Epitope Mapping Process
Sample
- Biosimilar
- Biobetter
- Antibody
- Protein-protein interaction
Analysis Process
- Ag-Ab complex
- 1) DEPC labeling
2) Chemical cross linking - Protease Digestion
- LC-MS
LC-MS/MS - Epitope Map
- Antigen
- Protease Digestion
- Immuno-affinity
biding assay - LC-MS
LC-MS/MS - Epitope Map
Analysis Result
1) DEPC labeling
2) Chemical cross link
3) Immuno-affinity peptide screening
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| Unbinding fraction | Elution fraction (Area) | sequence | ||||
|---|---|---|---|---|---|---|
| Frag# | Res | Area | 1st | 2nd | 3rd | |
| T14 | 180-188 | 1,291,000,000 | 810,600 | 8,799,000 | 11,950,000 | TAAGGPVTSR |
| T16 | 235-243 | 799,700,000 | 2,995,500 | 3,023,000 | 2,819,000 | VDDPPQNGDNDR |
| T21 | 281-288 | 66,900,000 | 1,425,000 | 1,232,000 | 1,367,000 | WQCQLYQGER |
Conclusion
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