This paper describes a versatile sensing membrane for immobilizationof proteins (antibodies, antigens, enzymes and so on) and a new immobolizationmethod' The membrane was developed with polyacrylaminde gel and blo...This paper describes a versatile sensing membrane for immobilizationof proteins (antibodies, antigens, enzymes and so on) and a new immobolizationmethod' The membrane was developed with polyacrylaminde gel and blotinstrep-tavidin system. Streptavidin was bond to the polyacryamide gel with 1-ethyl-3-(3-dimethylaminopropyl carbodimide (EDAC), which results in a streptavidin membrane, and a biotinalyted ligand was then easily linked to the gel via the streptavidin molecules.The streptavidin membrane (the thickness or 500urn) was used toimmobilize anti-human chorionic gonadotrophin antibody (Q hCG). The number orQbCG in the membrane was about 1011 per mm2 of the membrane surface. The membrane was used as a sensing membrane or optical immunosensor (Mach-Zehnder interferomter). It had a good response at the concentration or 10 afl/L hCG.展开更多
A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immob...A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3',5,5'-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.展开更多
文摘This paper describes a versatile sensing membrane for immobilizationof proteins (antibodies, antigens, enzymes and so on) and a new immobolizationmethod' The membrane was developed with polyacrylaminde gel and blotinstrep-tavidin system. Streptavidin was bond to the polyacryamide gel with 1-ethyl-3-(3-dimethylaminopropyl carbodimide (EDAC), which results in a streptavidin membrane, and a biotinalyted ligand was then easily linked to the gel via the streptavidin molecules.The streptavidin membrane (the thickness or 500urn) was used toimmobilize anti-human chorionic gonadotrophin antibody (Q hCG). The number orQbCG in the membrane was about 1011 per mm2 of the membrane surface. The membrane was used as a sensing membrane or optical immunosensor (Mach-Zehnder interferomter). It had a good response at the concentration or 10 afl/L hCG.
基金supported by the National Natural Science Foundation of China (30825027)Special Fund for Agro-Scientific Research in the Public Interest (200903009)
文摘A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3',5,5'-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.
