Figure 1.

Experimental strategy and identified proteins from the discovery experiment for ALS biomarker candidate proteins. Human CSF samples from 20 patients with ALS and 20 HC individuals were analyzed with four 11-plex TMT experimental batches. MP was prepared by pooling all 40 CSF samples and was added to each TMT experiment to normalize between batches. CSF proteins were digested with trypsin and labeled with TMT reagents, followed by pre-fractionation into 24 fractions prior to mass spectrometry analysis. Proteins were subsequently identified by conducting a database search of the acquired mass spectra (A). The numbers of identified proteins from 4 batches of TMT-based proteomics experiments are shown in the Venn diagram (B).