Enzyme-Linked Immunosorbent Assay-Based Microarray on a Chip for Bioaerosol Sensing: Toward Sensitive and Multiplexed Profiling of Foodborne Allergens

Food allergy has become a growing health concern that may impair life quality and even cause life-threatening outcomes. Accidental and continuous exposure to allergenic bioaerosols has a substantially negative impact on the respiratory health of patients. Traditional analytical methodologies for food allergens are restricted by strong reliance on bulk instrumentation and skilled personnel, particularly in low-resource settings. In this study, a fluorescent sensor array based on the enzyme-linked immunosorbent assay performed on a herringbone-shaped microfluidic chip (ELISA-HB-chip) was designed for dynamically sensitive and multiplexed quantification of foodborne allergens in aerosols that originated from liquid food extracts. Due to the high surface area of aerosol particles and sufficient mixing of immunological reagents using a herringbone micromixer, the detection sensitivity was improved by over an order of magnitude compared to traditional allergen detection in the aqueous phase. Through fluorescence imaging of multiple regions on the ELISA-HB-chip, four important foodborne allergens, namely, ovalbumin, ovomucoid, lysozyme, and tropomyosin, could be simultaneously monitored without any cross-reactivity, and the limits of detection for these allergenic species were determined to be 7.8, 1.2, 4.2, and 0.31 ng/mL, respectively. Combining with a 3D printed and portable fluorescence microscope, this platform exhibited an excellent field-deployable capacity for quick and accurate determination of allergens in the aerosol state from spiked buffer solutions, thus displaying the practicality for food safety screening at cooking or food processing sites where patients are potentially under exposure to allergenic bioaerosols that escaped from food matrices or extracts.