Comparison of non-magnetic and magnetic beads in bead-based assays

Silvie Hansenová Maňásková, Alex van Belkum, Hubert Endtz, Floris J. Bikker, Enno C.I. Veerman, Willem van Wamel

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11 Citations (Scopus)

Abstract

Multiplex bead-based flow cytometry is an attractive way for simultaneous, rapid and cost-effective analysis of multiple analytes in a single sample. Previously, we developed various bead-based assays using non-magnetic beads coated with Staphylococcus aureus and Streptococcus pneumoniae antigens for the detection of antibodies. Here, we compared the performance of the assay using non-magnetic beads with one based on the newly developed magnetic beads. We optimized the magnetic beads' coupling procedure and antibody detection assays for S. aureus and S. pneumoniae antigens and we measured IgG in human pooled serum against a series of S. aureus and S. pneumoniae-derived antigens in a singleplex and in a multiplex assay, respectively. For the multiplex assay, the comparison between magnetic and non-magnetic beads showed: i) in the majority of the cases (13 of the 17 tested S. pneumoniae antigens) significantly higher Median Fluorescence Intensity (ME) values, ii) lower detection limits, iii) lower coefficient of variation (CV: 12% vs. 7% for non-magnetic vs. magnetic beads), so lower inter assay variation and hence higher reproducibility. Magnetic bead coupling is cost effective, as we used 25% of the normal amount of antigen and only 50% of the beads in comparison to the non-magnetic beads. This optimized magnetic-based assay, which combines ease of use with an improved assay performance, allows detection of antibodies with a low titer that are potentially missed with the non-magnetic-based assay. (C) 2016 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)29-33
Number of pages5
JournalJournal of Immunological Methods
Volume436
DOIs
Publication statusPublished - Sept 2016

Bibliographical note

© 2016 Elsevier B.V. All rights reserved.

This work was supported by Sanofi Pasteur; grant title: Determination of childhood pneumonia associated with Streptococcus pneumoniae with the project number: PR-10062 and by ZonMW; grant title: Exploiting Staphylococcus aureus sortase for anti-infective purposes with the project number: 50-51700-98-055

Research programs

  • EMC MM-04-28-01

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