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A hydrogel sensor-based microfluidic platform for the quantitative and multiplexed detection of fertility markers for point-of-care immunoassays
- Kalme, Satish, Kandaswamy, Srinivasan, Chandrasekharmath, Anusha, Katiyar, Reeta, Rajamanickam, Gokul Prasath, Kumar, Suraj, Dendukuri, Dhananjaya
- Analytical methods 2019 v.11 no.12 pp. 1639-1650
- analytical methods, antibodies, automation, buffers, cross reaction, follicle-stimulating hormone, hydrogels, immunoassays, in vitro fertilization, luteinization, luteinizing hormone, manufacturing, organ-on-a-chip, point-of-care systems, prolactin
- We report a new microfluidics-based immunoassay platform based on 3D porous hydrogel particle sensors with planar rectangular geometries. Large numbers of sensors are made off-chip before being embedded into an injection molded microfluidic chip. This allows for both multiplexing and scalability of manufacturing processes. A specially designed adapter containing detection reagents and buffers is interfaced with a microfluidic chip allowing for an automated immunoassay to be performed. All antibody reagents required for the test are stored in a dried form on the chip, enabling stability for resource-limited settings. Fluorescence-based immunoassays are performed by a sequential flow of the sample, detection reagents and wash buffer over the sensors. A compact instrument that integrates microfluidic flow dispensing and pumping with highly sensitive array-based detection is used to conduct and read the assay. A multiplexed set of 4 important fertility-linked tests, beta-human chorionic gonadotropin (β-hCG), follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL), that are commonly required at gynecology/IVF settings were developed and validated using discarded clinical samples. Detailed data on accuracy, precision, linearity, effects of interference and cross-reactivity have been studied, rendering the platform ready for clinical use.