Abstract
Protein analysis is the foundation to understanding the mechanisms of complex biological processes. As one of the most widely used techniques to determine protein species and contents, protein dot blot aids biology research but needs corresponding antibodies for marking. A label-free detection method based on terahertz time-domain spectroscopy (THz-TDS) is proposed and demonstrated to improve this traditional technology. A membrane loaded with protein samples is directly scanned using a transmission THz-TDS system for spectral imaging. Different kinds of proteins can be distinguished by the refractive index extracted from the THz transmission spectrum. The intensity or shade imaged with the THz transmission spectrum can help detect the protein quantitatively. The feasibility of this new protein assay is demonstrated by the results of systematic testing with actual samples prepared with the dot-blot protocol.
Highlights
Proteins play an important role within all living organisms
The quoted refractive index in the curves is the average of the refractive index of 27 single points among which every 9 were chosen at random from every 3 vertically adjacent protein sample points with the same protein quantity
A label-free method to acquire the quantitative distribution of different kinds of proteins using THz time-domain spectroscopy (THz-TDS) is presented
Summary
Proteins play an important role within all living organisms. As enzyme [1], antibody [2], signaling molecules [3], transporting molecules [4], and so on, they belong to a category of organic macromolecules participating in the formation of cells, and are principally responsible for life activities in the organism. The protein immunoblot as the most significant semi quantitative experimental analysis method is conventionally used in the research on the expression of gene at the protein level, detection of the activity of antibody and early diagnosis of disease, due to its high specificity and sensitivity in solid phase immunoassay [12]. It takes advantages of the specific expression information of homologous antibody of the protein to determinate the presence or absence of the specific protein, together with its quantity. A reliable and label-free protein analysis method would help improve the detection efficiency and save time and money
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