title: MALDI-target integrated platform for affinity-captured protein digestion.
abstract: To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, ISET (integrated selective enrichment target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ranging from 40 fmol to 1 pmol per nanovial. On-ISET digestion and MALDI MS/MS analysis of immunoaffinity captured disease-associated biomarker PSA (prostate specific antigen) from human seminal plasma are presented.
authors.affiliation: Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Box 118, 22100 Lund, Sweden; CREATE Health, Lund University, Medicon Village, Bldn 406, 22381 Lund, Sweden; Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, Serdang, Selangor 43400, Malaysia.
authors.forename: Asilah
authors.initials: A
authors.lastname: Ahmad-Tajudin
authors.affiliation: Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Box 118, 22100 Lund, Sweden; CREATE Health, Lund University, Medicon Village, Bldn 406, 22381 Lund, Sweden. Electronic address: Belinda.Adler@elmat.lth.se.
authors.forename: Belinda
authors.initials: B
authors.lastname: Adler
authors.affiliation: Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Box 118, 22100 Lund, Sweden; CREATE Health, Lund University, Medicon Village, Bldn 406, 22381 Lund, Sweden.
authors.forename: Simon
authors.initials: S
authors.lastname: Ekström
authors.affiliation: Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Box 118, 22100 Lund, Sweden.
authors.forename: György
authors.initials: G
authors.lastname: Marko-Varga
authors.affiliation: Department of Laboratory Medicine, Division of Clinical Chemistry, Lund University, Skåne University Hospital, 20502 Skåne, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Malm
authors.affiliation: Department of Laboratory Medicine, Division of Clinical Chemistry, Lund University, Skåne University Hospital, 20502 Skåne, Sweden; Department of Laboratory Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA; Department of Surgery (Urology Service), Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA; Department of Medicine (GU Oncology Service), Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA; Nuffield Department of Surgical Sciences, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom; Institute of Biomedical Technology, University of Tampere, Tampere 33520, Finland.
authors.forename: Hans
authors.initials: H
authors.lastname: Lilja
authors.affiliation: Department of Measurement Technology and Industrial Electrical Engineering, Lund University, Box 118, 22100 Lund, Sweden; CREATE Health, Lund University, Medicon Village, Bldn 406, 22381 Lund, Sweden; Department of Biomedical Engineering, Dongguk University, Seoul, Republic of Korea.
authors.forename: Thomas
authors.initials: T
authors.lastname: Laurell
mesh_terms: D000478:Alkylation; D000911:Antibodies, Monoclonal / Q000276:immunology; D002846:Chromatography, Affinity; D006801:Humans; D008297:Male; D010084:Oxidation-Reduction; D017430:Prostate-Specific Antigen / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D040901:Proteomics; D052616:Solid Phase Extraction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000911:Antibodies, Monoclonal; D017430:Prostate-Specific Antigen
keywords: Digestion; ISET; Immunoaffinity; MALDI; MRM; MS; PSA; Proteomics; RP; SPE; SPR; integrated selective enrichment target; mass spectrometry; matrix-assisted laser desorption/ionization; multiple reaction monitoring; prostate specific antigen; reverse phase; solid-phase extraction; surface plasmon resonance