The advantages, challenges, and applications of each target protein quantification technique.
Advantages
Challenges
Applications
SRM/MRM-based assays
(i) High selectivity (ii) Low development cost (iii) Can be developed rapidly (iv) Multiplex possibility (v) Distinguish close related targets (vi) Measure low or nonimmunogenic targets
(i) Low sensitivity (ii) Not easy to standardize (multiple efforts in progress) (iii) Complex sample preparation (iv) High instrument cost (v) Relatively low throughput
(i) Relatively high abundant targets without antibody for initial analytical validation (ii) Small peptide targets (iii) Low or nonimmunogenic targets
ELISA, immunoassays
(i) High sensitivity (ii) Low operating cost (iii) No or simple sample preparation (iv) Easy to standardize and distribute (v) High throughput (vi) Automation
(i) Development cost is high (ii) Long timeframe to acquire a good antibody (iii) Hook-effects (iv) Highly challenging on PTM (v) Very hard to deal with low-immunogenic target (vi) Cross-reactivity
(i) Clinical applications when good reagents are available (ii) Low abundant targets
Immuno-mass spectrometry
(i) High selectivity (ii) High sensitivity (iii) Multiplex possibility (iv) Distinguish close related targets
(i) Still requires at least one high-affinity antibody and an expensive instrumentation
(i) Low abundant target and at least one high affinity antibody available (ii) Undistinguishable by antibody
