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ELISA (enzyme-linked immuno sorbent assay) method based on pH (potential of hydrogen) meter

An enzyme-linked immunosorbent assay technology, which is applied in the field of enzyme-linked immunosorbent assay based on pH meter, can solve the problems such as inability to use on-site analysis and instant inspection, high analysis cost, and large volume, and achieves reduction of analysis cost. Application prospect and effect of improving detection sensitivity

Active Publication Date: 2015-08-19
GUILIN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these optical instruments are expensive and bulky, resulting in high analysis costs of traditional ELISA methods, and cannot be used for on-site analysis and point-of-care testing.

Method used

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  • ELISA (enzyme-linked immuno sorbent assay) method based on pH (potential of hydrogen) meter
  • ELISA (enzyme-linked immuno sorbent assay) method based on pH (potential of hydrogen) meter
  • ELISA (enzyme-linked immuno sorbent assay) method based on pH (potential of hydrogen) meter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 40 pg / mL human oncogene protein (HOP) p190 / bcr-abl antigen was detected using a pH meter-based ELISA method.

[0020] The specific implementation process is as follows:

[0021] Such as figure 1 As shown, the specific steps of this example are: Step 1, add 50 μL 0.25 mg / mL human oncogene protein (HOP) p190 / bcr-abl monoclonal antibody (10 mM carbonic acid Sodium bicarbonate-sodium carbonate buffer solution, pH 9.6), let stand overnight at 4°C in the refrigerator, discard the liquid, spin dry, fill each well with washing solution (10 mM sodium dihydrogen phosphate-dipotassium hydrogen phosphate buffer solution , pH 7.4), let it stand for 30 s and discard it, repeat this 5 times, invert the well on a flat roll paper and pat dry; step 2, add 50 μL 10 mg / mL bovine serum white to a single enzyme-labeled well Protein (BSA) (prepared in 10 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.6), let stand at room temperature for 5 h, discard the liquid, shake dry, the...

Embodiment 2

[0024] The human oncogene protein (HOP) p190 / bcr-abl antigen was detected at concentrations ranging from 0.62 to 40 pg / mL using a pH meter-based ELISA method.

[0025] The specific implementation process is as follows:

[0026] Such as figure 1As shown, the specific steps of this example are: Step 1, add 50 μL 0.25 mg / mL human oncogene protein (HOP) p190 / bcr-abl monoclonal antibody (10 mM carbonic acid Sodium bicarbonate-sodium carbonate buffer solution, pH 9.6), let stand overnight at 4°C in the refrigerator, discard the liquid, spin dry, fill each well with washing solution (10 mM sodium dihydrogen phosphate-dipotassium hydrogen phosphate buffer solution , pH 7.4), let it stand for 30 s and discard it, repeat this 5 times, invert the well on a flat roll paper and pat dry; step 2, add 50 μL 10 mg / mL bovine serum white to a single enzyme-labeled well Protein (BSA) (prepared in 10 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.6), let stand at room temperature f...

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Abstract

The invention discloses an ELISA (enzyme-linked immuno sorbent assay) method based on a pH (potential of hydrogen) meter. The ELISA method comprises the following steps of performing antigen-antibody specificity immune reaction in an ELISA plate, capturing the antigen or antibody of the analyzing matter in a sample, introducing the glucose oxidase-glucose biological catalytic reaction, and using the pH meter to measure the pH value of the mixed product solution containing the glucose, wherein the size of the pH value is negatively related with the concentration of the analyzing matter in the sample, so as to complete the ELISA based on the pH meter. The ELISA method has the advantages that the signal is read by the low cost pH meter, so the cost of quantitative analysis is reduced; the response signal of single antigen-antibody bonding reaction is amplified by the micrometer or nanometer immobilized glucose oxidase; while the good specificity of the traditional ELISA method is maintained, the immobilizing amount of the glucose oxidase is increased by the micrometer or nanometer particles; the response signal of the single antigen-antibody specificity bonding reaction is amplified, and the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the technical field of enzyme-linked immunosorbent assay (Enzyme-Linked Immune-Sorbent Assay, abbreviated as ELISA), and specifically relates to an enzyme-linked immunosorbent assay method based on a pH meter. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) is an experimental technique with high specificity and high sensitivity, which is based on immunological reactions and combines the specific reactions of antibodies and antigens with the high-efficiency catalysis of enzymes on substrates. According to the different types and properties of the substances to be tested in the sample, various types of ELISA methods can be designed, including antibody sandwich method, antigen sandwich method, competition method, indirect method and capture coating method, etc. Since Engvall and Perlmann of Stockholm University in Sweden first established the enzyme-linked immunosorbent assay method in 1971, after more than...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 张云聂瑾芳杨佳妮范金龙高东
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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