Abstract
Serial measurement of a large panel of protein biomarkers near the bedside could provide a promising pathway to transform the critical care of acutely ill patients. However, attaining the combination of high sensitivity and multiplexity with a short assay turnaround poses a formidable technological challenge. Here, the authors develop a rapid, accurate, and highly multiplexed microfluidic digital immunoassay by incorporating machine learning-based autonomous image analysis. The assay has achieved 12-plexed biomarker detection in sample volume <15 μL at concentrations < 5 pg/mL while only requiring a 5-min assay incubation, allowing for all processes from sampling to result to be completed within 40 min. The assay procedure applies both a spatial-spectral microfluidic encoding scheme and an image data analysis algorithm based on machine learning with a convolutional neural network (CNN) for pre-equilibrated single-molecule protein digital counting. This unique approach remarkably reduces errors facing the high-capacity multiplexing of digital immunoassay at low protein concentrations. Longitudinal data obtained for a panel of 12 serum cytokines in human patients receiving chimeric antigen receptor-T (CAR-T) cell therapy reveals the powerful biomarker profiling capability. The assay could also be deployed for near-real-time immune status monitoring of critically ill COVID-19 patients developing cytokine storm syndrome.
Keywords: CAR-T therapy; Cytokine release syndrome; Machine learning; Microfluidic digital immunoassay; Multiplex biomarker detection.
【저자키워드】 machine learning, Cytokine release syndrome, CAR-T therapy, Microfluidic digital immunoassay, Multiplex biomarker detection., 【초록키워드】 therapy, Biomarker, Cytokines, Critical care, Cell therapy, machine learning, CAR-T, Cytokine storm syndrome, Image analysis, Antigen, Cytokine release syndrome, CNN, Protein, immunoassay, sensitivity, Critically ill, Algorithm, Convolutional neural network, Patient, Data analysis, pathway, sampling, longitudinal, chimeric antigen receptor, Immune status, Combination, Concentration, Analysis, COVID-19 patient, CAR-T therapy, critically ill COVID-19 patients, chimeric, Volume, serum cytokines, Longitudinal data, serum cytokine, machine, acutely ill patients, approach, Cell, develop, receiving, unique, reduce, reveal, autonomous, protein concentrations, 【제목키워드】 cytokine, immunoassay, Microarray, Analysis, machine,