title: Selective enrichment and sensitive detection of peptide and protein biomarkers in human serum using polymeric reverse micelles and MALDI-MS.
abstract: Reverse-micelle forming amphiphilic homopolymers with carboxylic acid and quaternary amine substituents are used to selectively enrich biomarker peptides and protein fragments from human serum prior to matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis. After depletion of human serum albumin (HSA) and immunoglobulin G (IgG), low abundance peptide biomarkers can be selectively enriched and detected by MALDI-MS at clinically relevant concentrations by using the appropriate homopolymer(s) and extraction pH value(s). Three breast cancer peptide biomarkers, bradykinin, C4a, and ITIH(4), were chosen to test this new approach, and detection limits of 0.5 ng mL(-1), 0.08 ng mL(-1), and 0.2 ng mL(-1), respectively, were obtained. In addition, the amphiphilic homopolymers were used to detect prostate specific antigen (PSA) at concentrations as low as 0.5 ng mL(-1) by targeting a surrogate peptide fragment of this protein biomarker. Selective enrichment and sensitive MS detection of low abundance peptide/protein biomarkers by these polymeric reverse micelles should be a sensitive and straightforward approach for biomarker screening in human serum.
authors.affiliation: Department of Chemistry, University of Massachusetts, LGRT 104 710 N. Pleasant St, Amherst, Massachusetts MA 01003, USA.
authors.forename: Nadnudda
authors.initials: N
authors.lastname: Rodthongkum
authors.forename: Rajasekhar
authors.initials: R
authors.lastname: Ramireddy
authors.forename: S
authors.initials: S
authors.lastname: Thayumanavan
authors.forename: W Vachet
authors.initials: WV
authors.lastname: Richard
mesh_terms: D015415:Biomarkers / Q000097:blood / Q000737:chemistry; D001798:Blood Proteins / Q000032:analysis* / Q000737:chemistry; D001920:Bradykinin / Q000097:blood / Q000737:chemistry; D002264:Carboxylic Acids / Q000737:chemistry; D015934:Complement C4a / Q000032:analysis* / Q000737:chemistry; D005260:Female; D006023:Glycoproteins / Q000097:blood / Q000737:chemistry; D006801:Humans; D008297:Male; D008823:Micelles; D010446:Peptide Fragments / Q000097:blood* / Q000737:chemistry; D017430:Prostate-Specific Antigen / Q000097:blood* / Q000737:chemistry; D053491:Proteinase Inhibitory Proteins, Secretory / Q000097:blood / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013501:Surface-Active Agents / Q000737:chemistry
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D002264:Carboxylic Acids; D006023:Glycoproteins; C091236:ITIH4 protein, human; D008823:Micelles; D010446:Peptide Fragments; D053491:Proteinase Inhibitory Proteins, Secretory; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; D015934:Complement C4a; D017430:Prostate-Specific Antigen; D001920:Bradykinin