A sensitive and selective assay of proteins is proposed based on measuring the total internal-reflected resonance light scattering(TIR-RLS) signals produced on the water/tetrachloromethane(H_2O/CCl_4) interface. In an...A sensitive and selective assay of proteins is proposed based on measuring the total internal-reflected resonance light scattering(TIR-RLS) signals produced on the water/tetrachloromethane(H_2O/CCl_4) interface. In an aqueous medium with pH value in the range of 3.29—3.78, electrostatic attraction occurs between the negatively charged Evans Blue(EB) and positively charged proteins, forming hydrophobic ion associates and resulting in EB-protein adsorption on H_2O/CCl_4 interface. The presence of cetyltrimethylammonium bromide prompts this adsorption, resulting in strongly enhanced TIR-RLS signals. The intensity of the enhanced TIR-RLS at 360—370 nm was found to be proportional to the concentration of proteins. For bovine serum albumin and human serum albumin, the linear range of detection is 0.07—1.2 μg/mL and the limits of detection are 6.68 and 6.30 ng/mL(3σ), respectively, while for lysozyme, the linear range of detection is 0.06—1.0 μg/mL and the limit of detection is 6.0 ng/mL(3σ). The content of the total albumin in a human urine sample could be directly determined by using the standard addition method with a percent recovery of 97.6%—104.1%, and the RSD ranging from 1.9% to 4.2%.展开更多
文摘A sensitive and selective assay of proteins is proposed based on measuring the total internal-reflected resonance light scattering(TIR-RLS) signals produced on the water/tetrachloromethane(H_2O/CCl_4) interface. In an aqueous medium with pH value in the range of 3.29—3.78, electrostatic attraction occurs between the negatively charged Evans Blue(EB) and positively charged proteins, forming hydrophobic ion associates and resulting in EB-protein adsorption on H_2O/CCl_4 interface. The presence of cetyltrimethylammonium bromide prompts this adsorption, resulting in strongly enhanced TIR-RLS signals. The intensity of the enhanced TIR-RLS at 360—370 nm was found to be proportional to the concentration of proteins. For bovine serum albumin and human serum albumin, the linear range of detection is 0.07—1.2 μg/mL and the limits of detection are 6.68 and 6.30 ng/mL(3σ), respectively, while for lysozyme, the linear range of detection is 0.06—1.0 μg/mL and the limit of detection is 6.0 ng/mL(3σ). The content of the total albumin in a human urine sample could be directly determined by using the standard addition method with a percent recovery of 97.6%—104.1%, and the RSD ranging from 1.9% to 4.2%.
