Robust Experimental Designs for the Development of the N-terminal proB Type Natriuretic Peptide (NT-proBNP) Assay Using Europium Chelate

Abstract

The Natriuretic peptides (BNP -brain natriuretic peptide (BNP) and ProBNP are markers with high sensitivity and specificity for heart failure, regulated by intra cardiac pressure. N-terminal pro B type natriuretic peptide (NT-proBNP) is an inactive product of BNP metabolism with a long half-life. BNPs are clinically valuable tools for risk stratification in patients with heart failure. The currently available assays are costly, and we need affordable techniques to make it available to all patients in various economic strata. Lateral flow immunoassay incorporated with fluorescence can be a better tool for quantifying the level of NT-proBNP in blood samples. Developing a point-of-care quantification of NT-proBNP concentration is challenging because of its low levels in blood in normal, healthy individuals. The study aimed to reduce the assay development time and cost by developing a europium chelated-based fluorescence immunoassay using a full factorial design approach. First, the control factors were mapped, and the transfer function was generated using the full factorial design of the experiment (DOE) approach. The control factors were ranked using a Pareto chart. The main effects and interaction effects were determined and plotted. The full factorial DOE also aims to reduce product development time by arriving at an optimum design point.