Fig 1: ELISA validation of 35plex TMTpro-MS hitsELISA was performed to provide orthogonal validation of putative plasma and CSF hits determined by 35plex TMTpro-MS using an independent biofluid cohort. A) Concentration of chondroadherin (CHAD) in healthy control and sALS plasma. B) Concentration of muscle creatine kinase (CKM) in healthy control and sALS plasma. C) Concentration of serpin family A group 3 (SERPINA3) in healthy control and sALS CSF. p-values denoted *, p < 0.05; ***, p < 0.001, ns- not significant.
Fig 2: Overview of 35-plex TMTpro-MS methodology for plasma and differentially expressed proteins between control and sALS.Flow diagram of A) proteomic workflow used to determine proteomic differences in plasma from (1) control versus sporadic ALS (sALS). Plasma is (2) immune-depleted to remove abundant proteins, (3) enzymatically digested, (4) undergoes TMT-labelling, and (4) samples are pooled prior to (5) analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). Volcano plot showing differentially expressed plasma proteins between B) controls (n= 31) versus sALS (n= 40). Box plots of the 8 significant proteins showing log2 relative abundance in control compared to sALS plasma. Each dot represents relative protein abundance in a single sample and line inside each box represents the median. Whiskers reflect minimum and maximum values. Proteins are shown in order of highest to least significance and include, C) carbonic anhydrase 3 (CA3), D) myoglobin (MB), E) creatine kinase M-type (CKM), F) neuronal cell adhesion molecule (NRCAM), G) chondroadherin (CHAD), H) platelet-activating factor acetylhydrolase (PLA2G7), I) serpin B6 (SERPINB6), and J) neural cell adhesion molecule 1 (NCAM1). Benjamini-Hochberg corrected adjusted p-values denoted *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Supplier Page from Abcam for Human CKM ELISA Kit