Patent Document ID: 10126309
Application ID: 14799778
Patent Flag: 1

Claim One:
1. A sample fractionation method for albumin biomarker identification comprising: (a) fractionating a sample derived from blood by size exclusion chromatography, wherein the sample has protein/peptide complexes dissociated by denaturation and the sample is spiked with a labeled protein of about 200 amino acid residues for differentiating protein and peptide pools; (b) collecting a (i) protein pool comprising the fractions preceding the end of the labeled protein peak, and (ii) a peptide pool comprising the fractions after the end of the labeled protein peak and before the free dye peak; (c) incubating the collected protein pool to allow for renaturation of the collected proteins in the protein pool; (d) depleting the protein pool by exposing the protein pool to an antibody depletion column and collecting column flow through to produce a depleted sample; (e) labeling the depleted sample with an uncharged thiol reactive label forming a saturated fluorescence labeled sample; (f) conducting two dimensional (2D) gel electrophoresis on the saturated fluorescence labeled sample, producing a 2D gel; (g) imaging the 2D gel and identifying a protein(s) of interest, wherein the protein of interest is an albumin isoform; and (h) conducting mass spectrometry analysis of the protein(s) of interest.