The instant invention provides an economical flow-through method for
determining amount of target proteins in a sample. An antibody
preparation (whether polyclonal or monoclonal, or any equivalent specific
binding agent) is used to capture and thus enrich a specific monitor
peptide (a specific peptide fragment of a protein to be quantitated in a
proteolytic digest of a complex protein sample) and an internal standard
peptide (the same chemical structure but including stable isotope
labels). Upon elution into a suitable mass spectrometer, the natural
(sample derived) and internal standard (isotope labeled) peptides are
quantitated, and their measured abundance ratio used to calculate the
abundance of the monitor peptide, and its parent protein, in the initial
sample.