Combination of abundant protein depletion and multi-lectin affinity chromatography (M-LAC) for plasma protein biomarker discovery.
We report on the development of a robust and relatively high-throughput method for in-depth proteomic analysis of human plasma suitable for biomarker discovery. The method consists of depletion of albumin and IgG and multi-lectin affinity chromatography (M-LAC), followed by nanoLC-MS/MS analysis of digested proteins and label-free comparative quantitation of proteins. The performance of the method is monitored by multiple quality control points to ensure reproducibility of the analysis. The method identifies proteins that are reported to be present in normal plasma at concentrations of 10-100 ng/mL and that may be of particular interest when studying a variety of disease conditions. Numerous tissue leakage proteins of potentially even lower concentrations are also identified. When the method was used in a study to identify potential biomarkers of psoriasis, the differential abundance of proteins present at low mug/mL level was quantitated and later verified by ELISA measurements.
Plavina
Tatiana
T
Biogen Idec Inc., Boston, MA 02142, USA.
Wakshull
Eric
E
Hancock
William S
WS
Hincapie
Marina
M
D000595:Amino Acid Sequence; D015415:Biomarkers / Q000097:blood*; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004798:Enzymes / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D037102:Lectins; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics; D011565:Psoriasis / Q000097:blood*; D012016:Reference Values; D017386:Sequence Homology, Amino Acid; D014357:Trypsin
D015415:Biomarkers; D001798:Blood Proteins; D004798:Enzymes; D037102:Lectins; D010446:Peptide Fragments; D014357:Trypsin