Filter Plate–Based Screening of MIP SPE Materials for Capture of the Biomarker Pro-Gastrin-Releasing Peptide. (December 2017)

• Record Type: Journal Article
• Authors: Jagadeesan, Kishore Kumar; Rossetti, Cecilia; Abdel Qader, Abed; Reubsaet, Léon; Sellergren, Börje; Laurell, Thomas; Ekström, Simon
• Is Part Of: SLAS discovery. Volume 22:Number 10(2017)
• Page Start: 1253
• Page End: 1261
• Publication Date: 2017-12
• Subjects: sample preparation -- mass spectrometry -- separations or chromatography -- protein chemistry -- protein labeling -- proteomics -- biomarkers; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; Biomolecules -- Analysis; Drugs -- Analysis; Drugs -- Testing; Drug Evaluation, Preclinical; Molecular Biology -- methods; Periodicals; Periodicals; 615.1
• DOI: 10.1177/2472555216689494
• Languages: English
• ISSNs: 2472-5552

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.