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Comparison of multiplex immunoassay platforms.
Clin Chem. 2010 Feb; 56(2):314-8.CC

Abstract

BACKGROUND

Candidate biomarkers discovered with high-throughput proteomic techniques (along with many biomarkers reported in the literature) must be rigorously validated. The simultaneous quantitative assessment of multiple potential biomarkers across large cohorts presents a major challenge to the field. Multiplex immunoassays represent a promising solution, with the potential to provide quantitative data via parallel analyses. These assays also require substantially less sample and reagents than the traditional ELISA (which is further limited by its ability to measure only a single antigen). We have measured the reproducibility, reliability, robustness, accuracy, and throughput of commercially available multiplex immunoassays to ascertain their suitability for serum biomarker analysis and validation.

METHODS

Assay platforms MULTI-ARRAY (Meso Scale Discovery), Bio-Plex (Bio-Rad Laboratories), A(2) (Beckman Coulter), FAST Quant (Whatman Schleicher & Schuell BioScience), and FlowCytomix (Bender MedSystems) were selected as representative examples of technologies currently used for high-throughput immunoanalysis. All assays were performed according to protocols specified by the manufacturers and with the reagents (diluents, calibrators, blocking reagents, and detecting-antibody mixtures) included with their kits.

RESULTS

The quantifiable interval determined for each assay and antigen was based on precision (CV < 25%) and percentage recovery (measured concentration within 20% of the actual concentration). The MULTI-ARRAY and Bio-Plex assays had the best performance with the lowest limits of detection, and the MULTI-ARRAY system had the most linear signal output over the widest concentration range (10(5) to 10(6)). Cytokine concentrations in unspiked and cytokine-spiked serum samples from healthy individuals were further investigated with the MULTI-ARRAY and Bio-Plex assays.

CONCLUSIONS

The MULTI-ARRAY and Bio-Plex multiplex immunoassay systems are the most suitable for biomarker analysis or quantification.

Authors+Show Affiliations

Departments of Medicine, Biomedical Engineering, Biological Chemistry, Bayview Proteomics Center, Johns Hopkins University, Baltimore, MD 21224, USA. qfu1@jhmi.eduNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20022982

Citation

Fu, Qin, et al. "Comparison of Multiplex Immunoassay Platforms." Clinical Chemistry, vol. 56, no. 2, 2010, pp. 314-8.
Fu Q, Zhu J, Van Eyk JE. Comparison of multiplex immunoassay platforms. Clin Chem. 2010;56(2):314-8.
Fu, Q., Zhu, J., & Van Eyk, J. E. (2010). Comparison of multiplex immunoassay platforms. Clinical Chemistry, 56(2), 314-8. https://doi.org/10.1373/clinchem.2009.135087
Fu Q, Zhu J, Van Eyk JE. Comparison of Multiplex Immunoassay Platforms. Clin Chem. 2010;56(2):314-8. PubMed PMID: 20022982.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of multiplex immunoassay platforms. AU - Fu,Qin, AU - Zhu,Jie, AU - Van Eyk,Jennifer E, Y1 - 2009/12/18/ PY - 2009/12/22/entrez PY - 2009/12/22/pubmed PY - 2010/3/5/medline SP - 314 EP - 8 JF - Clinical chemistry JO - Clin. Chem. VL - 56 IS - 2 N2 - BACKGROUND: Candidate biomarkers discovered with high-throughput proteomic techniques (along with many biomarkers reported in the literature) must be rigorously validated. The simultaneous quantitative assessment of multiple potential biomarkers across large cohorts presents a major challenge to the field. Multiplex immunoassays represent a promising solution, with the potential to provide quantitative data via parallel analyses. These assays also require substantially less sample and reagents than the traditional ELISA (which is further limited by its ability to measure only a single antigen). We have measured the reproducibility, reliability, robustness, accuracy, and throughput of commercially available multiplex immunoassays to ascertain their suitability for serum biomarker analysis and validation. METHODS: Assay platforms MULTI-ARRAY (Meso Scale Discovery), Bio-Plex (Bio-Rad Laboratories), A(2) (Beckman Coulter), FAST Quant (Whatman Schleicher & Schuell BioScience), and FlowCytomix (Bender MedSystems) were selected as representative examples of technologies currently used for high-throughput immunoanalysis. All assays were performed according to protocols specified by the manufacturers and with the reagents (diluents, calibrators, blocking reagents, and detecting-antibody mixtures) included with their kits. RESULTS: The quantifiable interval determined for each assay and antigen was based on precision (CV < 25%) and percentage recovery (measured concentration within 20% of the actual concentration). The MULTI-ARRAY and Bio-Plex assays had the best performance with the lowest limits of detection, and the MULTI-ARRAY system had the most linear signal output over the widest concentration range (10(5) to 10(6)). Cytokine concentrations in unspiked and cytokine-spiked serum samples from healthy individuals were further investigated with the MULTI-ARRAY and Bio-Plex assays. CONCLUSIONS: The MULTI-ARRAY and Bio-Plex multiplex immunoassay systems are the most suitable for biomarker analysis or quantification. SN - 1530-8561 UR - https://www.unboundmedicine.com/medline/citation/20022982/Comparison_of_multiplex_immunoassay_platforms_ L2 - https://academic.oup.com/clinchem/article-lookup/doi/10.1373/clinchem.2009.135087 DB - PRIME DP - Unbound Medicine ER -