Comparison of pre-processing methods for multiplex bead-based immunoassays

Tanja K Rausch, Arne Schillert, Andreas Ziegler, Angelika Lüking, Hans-Dieter Zucht, Peter Schulz-Knappe

Abstract

BACKGROUND: High throughput protein expression studies can be performed using bead-based protein immunoassays, such as the Luminex® xMAP® technology. Technical variability is inherent to these experiments and may lead to systematic bias and reduced power. To reduce technical variability, data pre-processing is performed. However, no recommendations exist for the pre-processing of Luminex® xMAP® data.

RESULTS: We compared 37 different data pre-processing combinations of transformation and normalization methods in 42 samples on 384 analytes obtained from a multiplex immunoassay based on the Luminex® xMAP® technology. We evaluated the performance of each pre-processing approach with 6 different performance criteria. Three performance criteria were plots. All plots were evaluated by 15 independent and blinded readers. Four different combinations of transformation and normalization methods performed well as pre-processing procedure for this bead-based protein immunoassay.

CONCLUSIONS: The following combinations of transformation and normalization were suitable for pre-processing Luminex® xMAP® data in this study: weighted Box-Cox followed by quantile or robust spline normalization (rsn), asinh transformation followed by loess normalization and Box-Cox followed by rsn.

Original languageEnglish
JournalBMC Genomics
Volume17
Issue number1
Pages (from-to)601
ISSN0888-7543
DOIs
Publication statusPublished - 11.08.2016

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