Unlock instant, AI-driven research and patent intelligence for your innovation.

An immunoassay method based on graphene oxide photoactivated horseradish peroxidase

A technology of horseradish peroxidase and immunoassay, which is applied in the field of nanomaterial technology and biological analysis and detection, and can solve the problems of unsatisfactory sensitivity

Active Publication Date: 2018-07-06
JIANGNAN UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this routine method is simple, the sensitivity of the result is not very satisfactory [Duffy S.L.; Murphy J.T.Biotechniques 2001,31,495-496,498,500-501; Roda A.; Simoni P.; Mirasoli M.et al.Anal.Bioanal.Chem. 2002, 372, 751-758]

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • An immunoassay method based on graphene oxide photoactivated horseradish peroxidase
  • An immunoassay method based on graphene oxide photoactivated horseradish peroxidase
  • An immunoassay method based on graphene oxide photoactivated horseradish peroxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] a. Preparation of graphene oxide nanomaterials: 0.5g graphite powder and 0.5g NaNO 3 Place in 16.5 mL of 98% H 2 SO 4 , kept stirring at room temperature for 24h; then kept in an ice bath for 10 minutes, 3g KMnO 4 Slowly added to the mixture, continued to stir for 1h; kept the temperature at 35°C and continued to stir for 50min, then added 40mL of deionized water; finally added 1400mL of deionized water and 30% H 2 o 2 to stop the reaction; the product was filtered, washed several times with 5% HCl and dried in an oven at 50°C;

[0019] b. Immobilizing the alpha-fetoprotein secondary antibody on the Au nanoparticles and the base sequence is 5'-SH-(CH 2 ) 6 - Capture DNA of AAAAAAAGAAGGAGGGGCGACT-3' 1 , forming Au NPs / Ab 2 / DNA 1 Complex, as a signal tag to induce detection signal generation; Au NPs / Ab 2 / DNA 1 The complex was prepared as follows: 100 mL of 0.01% HAuCl 4The solution was boiled under vigorous stirring, and then 2.5mL of 1% trisodium citrate sol...

Embodiment 2

[0022] a. Preparation of graphene oxide nanomaterials: 0.5g expanded graphite and 0.5g NaNO 3 Place in 16.5 mL of 98% H 2 SO 4 , kept stirring at room temperature for 24h; then kept in an ice bath for 10 minutes, 3g KMnO 4 Slowly added to the mixture, continued to stir for 1h; kept the temperature at 40°C and continued to stir for 30min, then added 40mL of deionized water; finally added 1400mL of deionized water and 30% H 2 o 2 to stop the reaction; the product was filtered, washed several times with 5% HCl and dried in an oven at 50°C;

[0023] b. Immobilizing the secondary antibody of alpha-fetoprotein on the Au nanoparticles and the base sequence is 5'-SH-(CH 2 ) 6 - Capture DNA of AAAAAAAGAAGGAGGGGCGACT-3' 1 , forming Au NPs / Ab 2 / DNA 1 Complex, as a signal tag to induce detection signal generation; Au NPs / Ab 2 / DNA 1 The complex was prepared as follows: 100 mL of 0.01% HAuCl 4 The solution was boiled under vigorous stirring, and then 2.5mL of 1% trisodium citra...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a graphene oxide photoactivation-based horseradish peroxidase (HRP) immunoassay method. Graphene oxide can generate an active intermediate photogenerated holes and superoxide anion free radicals under lights to realize catalytic oxidation of a chromogenic substrate by the HRP without H2O2. Alpha-fetoprotein (AFP) is adopted as a model, a DNA hybridization chain reaction is carried out to amplify a signal, and the HRP photoactivation effect of graphene oxide is used to realize catalytic oxidation of the substrate in order to realize immunodetection. The method combining the advantages of a nano-material and natural enzyme makes the detection limit of the AFP reach 0.0001 pg / mL. The method provided by the invention is a novel method for regulating the activity of the HRP through a photoexcited nano-material, and provides a new way for super sensitive enzyme-linked immunoassay.

Description

Technical field: [0001] The invention relates to the field of nanomaterial science and technology and the field of biological analysis and detection, in particular to light-induced enzyme catalysis based on nanomaterials and its application in immune analysis and detection. Background technique: [0002] Enzyme-linked immunosorbent assay (ELISA) is a new type of immunoassay technology developed on the basis of immunoenzyme technology, which is a frontier topic in the field of analytical chemistry. Its core technology is the combination of antigen or antibody and enzyme complex, and then generate signal through enzyme-catalyzed reaction to detect antigen or antibody [Kawatsu K.; Hamano Y.; SugiyamaA.et al.J.Food Protect.2002,65, 1304-1308; Micheli L.; Di Stefano S.; Moscone D. et al. Anal. Bioanal. Chem. 2002, 373, 678-684]. It has the advantages of rapidity, strong specificity and easy operation, etc. It has been widely used in various fields of chemistry and life sciences,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535B82Y30/00B82Y40/00
CPCB82Y30/00B82Y40/00G01N33/535
Inventor 王光丽曹根霞董玉明李小琴
Owner JIANGNAN UNIV