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.

MALDIViz: A Comprehensive Informatics Tool for MALDI-MS Data Visualization and Analysis

“…Briefly, six different SPE materials in duplicate were examined using four different SPE conditions in a micro-SPE platform. 17 Without dedicated analysis software such as MALDIViz, a user will have to compare these mass spectra manually, which is both tedious and very risky from an analytical perspective. The visualization methods, in combination with user-defined sample grouping by coloring, provided a fast and convenient means to identify the experimental trends.…”

“…Briefly, six different SPE phases in duplicate were examined using four different SPE conditions using a 384-plate-based micro-SPE platform. Thereafter, the collected eluents were analyzed in triplicate by MALDI-MS. 17…”

“…Briefly, six different SPE phases in duplicate were examined using four different SPE conditions using a 384-plate-based micro-SPE platform. Thereafter, the collected eluents were analyzed in triplicate by MALDI-MS. 17 Dataset 2: Cancer Dataset. This is a publicly available dataset from a MALDI-based sample profiling publication using gold nanoparticles in order to separate the proteins and peptides in human serum.…”

“…MALDIViz is an R-Shiny-based web application, accessible independently of the operating system and without the need to install the program locally. The functionality of MALDIViz application is demonstrated by analysis of in-house-generated screening data from an investigation of different solid-phase extraction (SPE) materials 17 and a publicly available cancer dataset. 18 The MALDIViz application is accessible through https://jkkishore85.shinyapps.io/maldiviz/.…”

Advances in MALDI Mass Spectrometry within Drug Discovery

“…Scientists are combining disparate technologies to create synergies with MALDI TOF. For example, Jagadeesan and coauthors 21 have applied molecular imprint technology for the capture and detection of progastrin-releasing peptide. Jagadeesan and Ekström 22 have further developed a visualization tool for MALDI data that can be quickly applied to the analysis of large datasets.…”