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Multidimensional Normalization to Minimize Plate Effects of Suspension Bead Array Data
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.ORCID iD: 0000-0001-8603-8293
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.ORCID iD: 0000-0002-4657-8532
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2016 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 15, no 10, p. 3473-3480Article in journal (Refereed) Published
Abstract [en]

Enhanced by the growing number of biobanks, biomarker studies can now be performed with reasonable statistical power by using large sets of samples. Antibody-based proteomics by means of suspension bead arrays offers one attractive approach to analyze serum, plasma, or CSF samples for such studies in microtiter plates. To expand measurements beyond single batches, with either 96 or 384 samples per plate, suitable normalization methods are required to minimize the variation between plates. Here we propose two normalization approaches utilizing MA coordinates. The multidimensional MA (multi-MA) and MA-loess both consider all samples of a microtiter plate per suspension bead array assay and thus do not require any external reference samples. We demonstrate the performance of the two MA normalization methods with data obtained from the analysis of 384 samples including both serum and plasma. Samples were randomized across 96-well sample plates, processed, and analyzed in assay plates, respectively. Using principal component analysis (PCA), we could show that plate-wise clusters found in the first two components were eliminated by multi-MA normalization as compared with other normalization methods. Furthermore, we studied the correlation profiles between random pairs of antibodies and found that both MA normalization methods substantially reduced the inflated correlation introduced by plate effects. Normalization approaches using multi-MA and MA-loess minimized batch effects arising from the analysis of several assay plates with antibody suspension bead arrays. In a simulated biomarker study, multi-MA restored associations lost due to plate effects. Our normalization approaches, which are available as R package MDimNornin, could also be useful in studies using other types of high-throughput assay data.

Place, publisher, year, edition, pages
2016. Vol. 15, no 10, p. 3473-3480
Keywords [en]
normalization, plate effect, affinity proteomics, multiplexed immunoassays
National Category
Chemical Sciences Engineering and Technology
Identifiers
URN: urn:nbn:se:kth:diva-196086DOI: 10.1021/acs.jproteome.5b01131ISI: 000385054100003PubMedID: 27570895Scopus ID: 2-s2.0-84990836997OAI: oai:DiVA.org:kth-196086DiVA, id: diva2:1047023
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QC 20161116

Available from: 2016-11-16 Created: 2016-11-11 Last updated: 2024-03-15Bibliographically approved

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Hong, Mun-GwanNilsson, PeterSchwenk, Jochen M.

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