Abstract
BackgroundProtein biomarker studies are currently hampered by a lack of measurement standards to demonstrate quality, reliability and comparability across multiple assay platforms. This is especially pertinent for immunoassays where multiple formats for detecting target analytes are commonly used.FindingsIn this pilot study a generic panel of six non-human protein standards (50 - 10^7 pg/mL) of varying abundance was prepared as a quality control (QC) material. Simulated "normal" and "diseased" panels of proteins were prepared in pooled human plasma and incorporated into immunoassays using the Meso Scale Discovery® (MSD®) platform to illustrate reliable detection of the component proteins. The protein panel was also evaluated as a spike-in material for a model immunoassay involving detection of ovarian cancer biomarkers within individual human plasma samples. Our selected platform could discriminate between two panels of the proteins exhibiting small differences in abundance. Across distinct experiments, all component proteins exhibited reproducible signal outputs in pooled human plasma. When individual donor samples were used, half the proteins produced signals independent of matrix effects. These proteins may serve as a generic indicator of platform reliability.Each of the remaining proteins exhibit differential signals across the distinct samples, indicative of sample matrix effects, with the three proteins following the same trend. This subset of proteins may be useful for characterising the degree of matrix effects associated with the sample which may impact on the reliability of quantifying target diagnostic biomarkers.ConclusionsWe have demonstrated the potential utility of this panel of standards to act as a generic QC tool for evaluating the reproducibility of the platform for protein biomarker detection independent of serum matrix effects.
Highlights
Protein biomarkers for diagnosis of disease have formed the basis of clinical research proteomics for several decades [1,2,3]
We have demonstrated the potential utility of this panel of standards to act as a generic quality control (QC) tool for evaluating the reproducibility of the platform for protein biomarker detection independent of serum matrix effects
The component proteins exhibiting differential signals between distinct samples would be indicative of the existence of matrix effects that may influence credibility in the data derived for the detection of the target analytes
Summary
Protein biomarkers for diagnosis of disease have formed the basis of clinical research proteomics for several decades [1,2,3]. Improved standardisation may be achieved through the use of generic protein standards, demonstrating the reproducibility of the platform function Such generic standards are emerging for mass spectrometry analysis of proteins, though they are specific to this platform rather than for broad stream applications including immunoassays [10]. Protein biomarker studies are currently hampered by a lack of measurement standards to demonstrate quality, reliability and comparability across multiple assay platforms. This is especially pertinent for immunoassays where multiple formats for detecting target analytes are commonly used
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