|
| Technique | 2DE (Two-dimensional gel electrophoresis) | SELDI-TOF MS | ICAT (Isotope-coded affinity tags) | IMAC (Immobilized metal affinity chromatography) |
|
| Principles | 2DE separates protein mixtures by
their isoelectric points and molecular weights, proteins can be identified by
MALDI-TOF MS through enzyme digestion | This technique employs protein
chip separates protein mixtures by different surface binding affinity and
molecular weights, and roughly identifies proteins through SELDI-TOF MS | ICAT separates proteins by
chemical labeling and relative abundance, and then obtains protein identification
through ESI MS/MS | IMAC is a powerful protein
fractionation method used to enrich metal-associated proteins and peptides,
proteins and peptides can be determined by ESI MS/MS |
|
| Remarks | Suitable for whole proteome or
specific pre-fractioned proteomes, detect large quantity of proteins in a single run,
not suitable for low abundant proteins, affected by posttranslational
modifications | Simple preparation procedures, sensitive detection limit,
small sample requirement, significant results,
wide detection range in molecular weight, modified by different surface
affinities | Biotinylated tags labeling before analysis;
suitable for low abundant proteins; not suitable for
post translational modified proteins;
more automated | Enrich metal-associated proteins
and peptides, easy regeneration, longevity and stability to proteolytic degradation,
have to be facilitated by other separation methods, suitable for posttranslational
modification |
|
| Potential applications | Study drug-induced cellular
signaling pathway in a global scale [35, 69]
| Identification of drug targets [70] | | |
| Identification of drug targets [70] | Screen drug candidates [72, 73]
| Receptome profiling [75, 76] | Mapping of phosphoproteomes [77, 78] and metalloproteomes [79, 80] |
| Study of drug toxicity and side
effects [71] | Study of protein-drug
interactions [74] | | |
|
