Label-free methods for performing assays using a colorimetric resonant optical biosensor
A technology of biosensors and reflective optics, applied in chemical instruments and methods, biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as high-throughput instruments that have not yet been produced
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Embodiment 1
[0202] Immobilized protein detection
[0203] To demonstrate the ability of the biosensor to quantify biomolecules on its surface, the 2 Droplets of BSA in O were applied to the biosensor shown in Figure 1. The 3 μl droplet was allowed to air dry, leaving a small amount of BSA distributed over a diameter of approximately 2 mm. The peak resonance wavelength at each biosensor location was measured before and after droplet deposition, and the peak wavelength shift was recorded. See Figure 34.
Embodiment 2
[0205] Immobilization of one or more specific binding substances
[0206] The following protocol was used on a colorimetric resonant reflectance biosensor to activate the surface with amine functional groups. Amine groups can be used as a universal surface for subsequent covalent attachment of several types of linker molecules.
[0207] Glass substrate biosensors of the present invention were cleaned by soaking in piranha etch (70 / 30% (v / v) concentrated sulfuric acid / / 30% hydrogen peroxide) for 12 hours. The biosensor was washed well with water. The biosensor was soaked in a solution of 3% 3-aminopropyltriethoxysilane in anhydrous acetone for 1 minute, then rinsed with anhydrous acetone, and air-dried. Alternatively, the biosensor was soaked in a solution of 10% 3-aminopropyltriethoxysilane (Pierce) in ethanol (Aldrich) for 1 min, followed by a brief ethanol rinse. The activated sensors were then dried at 70 °C for 10 min. The biosensor was then washed with water.
[0208...
Embodiment 3
[0219] IgG assay
[0220] As an initial demonstration of detection of biochemical binding, an assay was performed in which a biosensor was prepared by activation with the amino group surface chemistry described in Example 2, followed by attachment of a biotin linker molecule. The biotin linker is used to specifically interact with the streptavidin receptor molecule and efficiently attach it to the surface by contacting 50 μg / ml Concentration of the streptavidin solution is achieved for 2-4 hours. The streptavidin receptor is capable of binding any biotinylated protein on the biosensor surface. For this example, 3 μl of biotinylated anti-human IgG droplets in phosphate buffered saline (PBS) were deposited on 4 different locations on the biosensor surface at a concentration of 200 μg / ml. The solution was incubated on the biosensor for 30 minutes and then rinsed extensively with PBS. The peak resonance wavelengths at the four positions were measured after biotin activation, af...
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