Abstract
We evaluated optical pickup ELISA with an original microfluidic disk that contains eight radially arranged channels, which enable semi-automatic sample loading and washing. This disk-shaped chip composed of acrylic plates was fabricated by CO2 laser machining and capture antibodies were immobilized in the channels. After the immunoreaction with antigens and enzyme-linked secondary antibodies, an enzyme-catalyzed nanoaggregation of o-phenylenediamine was detected by measuring the reflectivity change of a laser beam focused in the channel. The assay of C-reactive protein (CRP) was successfully performed in a short amount of time (approximately 20 min from CRP loading). The limit of detection was determined to be 2 ng mL−1, which is more sensitive as compared with conventional ELISA using microplates.
Highlights
Bioanalyses with microfabricated devices, in which various microstructures for sample delivering, mixing, and signal detecting are integrated, have been actively studied in recent years.[1,2,3,4] Enzyme-linked immunosorbent assay (ELISA) is a powerful and commonly used technique to sensitively detect speci c proteins; recently, ELISA with microfabricated chips has attracted much attention
As an innovative way to solve the problems inherent in colorimetric-based ELISAs, we propose the application of optical pickup detection of horseradish peroxidase (HRP), which was recently developed by our group.[14,15]
Sophisticated disk-shaped chips have been designed and developed to minimize manual labour in ELISA, here we propose a disk-shaped chip for optical pickup ELISA with a simple design to semi-automate the washing process
Summary
Bioanalyses with microfabricated devices, in which various microstructures for sample delivering, mixing, and signal detecting are integrated, have been actively studied in recent years.[1,2,3,4] Enzyme-linked immunosorbent assay (ELISA) is a powerful and commonly used technique to sensitively detect speci c proteins; recently, ELISA with microfabricated chips (microELISA) has attracted much attention. Increased immobilization density and control of antibody orientation have been used by some research groups to enhance the efficiency of antigen–antibody interactions (immunoreactions) in small volumes.[7,8] While these approaches improve assay sensitivity, they are time-consuming and laborious in most cases. Another approach for signal improvement is the development of sensitive signal detection methods for low sample volumes
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