Abstract
A new fluorescent arrayed biosensor has been developed to discriminate species and concentrations of target proteins by using plural different phospholipid liposome species encapsulating fluorescent molecules, utilizing differences in permeation of the fluorescent molecules through the membrane to modulate liposome-target protein interactions. This approach proposes a basically new label-free fluorescent sensor, compared with the common technique of developed fluorescent array sensors with labeling. We have confirmed a high output intensity of fluorescence emission related to characteristics of the fluorescent molecules dependent on their concentrations when they leak from inside the liposomes through the perturbed lipid membrane. After taking an array image of the fluorescence emission from the sensor using a CMOS imager, the output intensities of the fluorescence were analyzed by a principal component analysis (PCA) statistical method. It is found from PCA plots that different protein species with several concentrations were successfully discriminated by using the different lipid membranes with high cumulative contribution ratio. We also confirmed that the accuracy of the discrimination by the array sensor with a single shot is higher than that of a single sensor with multiple shots.
Highlights
Important biomolecules such as DNA, RNA, proteins and so on are commonly detected by fluorescence techniques based on labeling and staining [1,2,3]
We have reported preliminary results on an arrayed biosensor utilizing liposomes to encapsulate fluorescent molecules and the time course analysis of the fluorescence [9]
We evaluated liposome-protein byanalysis investigating the change intensity of fluorescence emissionsthe using our developedinteraction software for
Summary
Important biomolecules such as DNA, RNA, proteins and so on are commonly detected by fluorescence techniques based on labeling and staining [1,2,3] Both the fluorescent antibody and the immunofluorescent technique are major techniques in the field of biochemistry because they offer high detectability, stability and safety. In the operation of the fluorescent liposome biosensor, the leakage of fluorescent molecules increases the fluorescence intensity It is further required: (1) to achieve a higher and more stable output intensity of fluorescence emission; (2) to investigate more different types of liposome phospholipid to improve the sensitivity and discrimination capability between different target biomolecules, and (3) to analyze the arrayed data statistically to increasing the accuracy of the analyzed results. Using PCA, we examined the feasibility and applicability of arrayed sensors with single shots in comparison with single microwell sensors with multiple shots
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