A Novel, Quantitative, Multi-analyte Immunoassay to Detect Biomarkers Involved in Lung Disease

M. Anderson, G. Hickey, I. O’Brien, P. Younge, L. Leong


Lung disorders affect 5–10% of the world’s population, and account for 1/6 of deaths worldwide. Pulmonary insult can arise from a multitude of factors, including those that are extrinsic, intrinsic, or iatrogenic-related. Manifestations of pulmonary disease include asthma, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), interstitial pneumonia (UIP), lung cancer, and tuberculosis (TB). The array of symptoms associated with these lung conditions involve shortness of breath, cough, chest pain, fever, pleural effusion, pleural thickening pulmonary fibrosis, necrobiotic nodules, and bronchiolitis obliterans organizing pneumonia (BOOP). A decline in the quality of life in patients with lung disease is experienced not only physically, but also mentally due to the severity of the symptoms. One example is lung cancer, which is the most common cause of death in developed countries. Patients diagnosed with stage III or IV lung cancer have a survival rate of only 15% with current therapies.

The study presented here looked at several cytokine markers involved in airway constriction including IL-13, IL-5, and IL-4 as well as select biomarkers known to be involved in the regulatory pathways for IFN-gamma and TNF-alpha. The Simple Plex™ assay that was employed is a novel, quantitative, multi-analyte immunoassay platform that delivers high precision and accuracy from only 25 μL of sample. This platform measures up to four analytes simultaneously from a single sample with very high sensitivity. It is a microfluidic-based system that allows parallel single analyte detection, and reduces non-specific antibody interactions often observed in other traditional multiplex platforms. The assay is a closed system, removing potential user variability. This is accomplished by enclosing the entire assay process within a single cartridge that provides results in just over one hour. Our preliminary studies suggest this platform may represent a highly efficient method for the quantitation of select biomarkers known to be involved in lung disease.