title: Filter Plate-Based Screening of MIP SPE Materials for Capture of the Biomarker Pro-Gastrin-Releasing Peptide.
abstract: Affinity-based solid-phase extraction (SPE) is an attractive low-cost sample preparation strategy for biomarker analysis. Molecularly imprinted polymers (MIPs) as affinity sorbents offer unique opportunities for affinity SPE, due to their low manufacturing cost and high robustness. A limitation is the prediction of their affinity; therefore, screening of analyte recovery and specificity within a large range of SPE conditions is important in order to ensure high-sensitivity detection and assay reproducibility. Here, a µ-SPE method for screening of the MIP-SPE materials using a commercial 384-well filter plate is presented. The method allows for rapid and automated screening using 10-30 µL of packed SPE sorbent per well and sample volumes in the range of 10-70 µL. This enables screening of many different SPE sorbents while simultaneously identifying optimal SPE conditions. In addition, the 384-well format also facilitates detection with a multitude of analytical platforms. Performance of the µ-MIP-SPE method was investigated using a series of MIPs designed to capture pro-gastrin-releasing peptide (ProGRP). Fractions coming from sample load, cartridge wash, and elution were collected and analyzed using mass spectrometry (MS). The top-performing MIPs were identified, together with proper SPE conditions.
authors.affiliation: 1 Department of Biomedical Engineering, Lund University, Lund, Sweden.
authors.forename: Kishore Kumar
authors.initials: KK
authors.lastname: Jagadeesan
authors.affiliation: 2 Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, Oslo, Norway.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Rossetti
authors.affiliation: 3 Department of Environmental Chemistry and Analytical Chemistry, Institute for Environmental Research (INFU), Technical University of Dortmund, Dortmund, Germany.
authors.forename: Abed
authors.initials: A
authors.lastname: Abdel Qader
authors.affiliation: 2 Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, Oslo, Norway.
authors.forename: Léon
authors.initials: L
authors.lastname: Reubsaet
authors.affiliation: 4 Department of Biomedical Sciences, Faculty of Health and Society, University of Malmö, Malmö, Sweden.
authors.forename: Börje
authors.initials: B
authors.lastname: Sellergren
authors.affiliation: 1 Department of Biomedical Engineering, Lund University, Lund, Sweden.
authors.forename: Thomas
authors.initials: T
authors.lastname: Laurell
authors.affiliation: 1 Department of Biomedical Engineering, Lund University, Lund, Sweden.
authors.forename: Simon
authors.initials: S
authors.lastname: Ekström
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D005374:Filtration / Q000295:instrumentation*; D019886:Gastrin-Releasing Peptide / Q000032:analysis*; D057166:High-Throughput Screening Assays; D054802:Molecular Imprinting / Q000379:methods*; D011108:Polymers / Q000737:chemistry; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D015415:Biomarkers; D011108:Polymers; D019886:Gastrin-Releasing Peptide
keywords: biomarkers; mass spectrometry; protein chemistry; protein labeling; proteomics; sample preparation; separations or chromatography