ABSTRACT
The coronavirus pandemic has profoundly changed the way medical education is delivered globally. Our group reports an insight into the adaptations and innovations made by the School of Medicine at Anglia Ruskin University.
ABSTRACT
BACKGROUND: 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. FINDINGS: In 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. CONCLUSIONS: We 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.
ABSTRACT
Cell surface protein profiling is a generic tool to determine the quality of cultured mammalian cell lines. Flow cytometry is commonly employed as the screening technique of choice, but it consumes a large quantity of cells. We have evaluated an electrochemiluminescent (ECL)-based platform, the Meso Scale Discovery (MSD) Sector 6000 Imager, for cell surface protein detection with fewer than 100 cells per well. The detection of CD13 on human foreskin fibroblast (HFF) cells was evaluated using indirect detection of the reverse-phase immunoassays in 96-well and 384-well plate formats. The study has shown robust cell surface detection of CD13 on HFF cells using the 384-well plate format, with a LOD of 15.3 ± 6.5 cells (n=3). The 96-well plate format exhibited far greater variability between experiments, such that the best LOD obtained was 42.5 cells per well. We have demonstrated, with our model system, the feasibility to perform cell-binding assays with as few as 15 cells per well of a 384-well microplate using the MSD platform.