Counting proteins without labels
The problem. To compare protein abundance across samples by mass spectrometry, the reliable route was chemical labelling (isotope tags), which is costly and limits sample number. Label-free quantification is cheaper and unlimited, but naive intensity comparison is noisy: run-to-run variation and missing values wreck the ratios you actually want.
The idea. MaxLFQ, built into MaxQuant, computes relative abundances from pairwise peptide ratios — extracting the maximum reliable ratio information across samples — and defers normalisation so it doesn’t distort those ratios. The result is label-free quantities accurate enough to stand in for labelled methods across large sample sets.
Why it matters. This is the proteomics analogue of the count-normalisation problem I know from RNA-seq: how do you make abundances comparable across runs of differing total signal? Seeing DESeq’s “model the systematic variation” instinct reappear in mass spec — different data, same discipline — is exactly the cross-modality pattern this reading list keeps surfacing. Proteomics has its own version of every RNA-seq problem.
Verdict. Foundational for quantitative proteomics and still the standard label-free approach. Read it right after MaxQuant (yesterday) — identification then quantification — and note how much it rhymes with the DE normalisation I already do on the RNA side.