Preparation method for artificial antibody for caffeine detection

An artificial antibody, caffeine technology, applied in the field of material science, can solve problems such as poor selectivity, and achieve the effect of easy operation, high sensitivity and high specificity

Active Publication Date: 2019-05-31
WUXI ZODOLABS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Chemiluminescence method has high sensitivity and wide linear range, but poor selectivity. Immunoassay is an analysis established by directly labeling luminescent substrates or enzymes on antigens or antibodies, and forming complexes through the specific combination of antigens and antibodies. method

Method used

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  • Preparation method for artificial antibody for caffeine detection
  • Preparation method for artificial antibody for caffeine detection
  • Preparation method for artificial antibody for caffeine detection

Examples

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preparation example Construction

[0056] A method for preparing an artificial antibody for caffeine detection, characterized in that: the artificial antibody elutes the imprinted molecules located in the polymer imprinted shell, and the inner part of the imprinted shell has the structure and size of the imprinted molecule With a hole structure complementary to the functional group, the artificial antibody that elutes the imprinted molecule has a specific recognition site for the enzyme-labeled antigen molecule, and realizes selective recognition of the enzyme-labeled antigen molecule. The luminescence method realizes trace detection of enzyme-labeled antigen molecules. The preparation process of the above-mentioned artificial antibody includes the following four steps:

[0057] The first step is the oxidation of horseradish peroxidase: Weigh 4 ~ 6mg of horseradish peroxidase and dissolve it in 1 ~ 3mL of freshly prepared acetate buffer solution containing sodium periodate, and react in the dark at room temperat...

specific Embodiment

[0061] Dissolving caffeine in acetonitrile solution reacts with functional monomer methacrylic acid, and the amino group and carbonyl group in the molecular structure of caffeine and the carboxyl group of methacrylic acid are arranged in the form of hydrogen bonds to form a hydrogen bond adduct. A cross-linking agent ethylene glycol dimethacrylate and an initiator azobisisobutyronitrile are added to the hydrogen bond adduct to obtain molecular polymer microspheres imprinted with caffeine. Using V (methanol): V (acetic acid) = 9: 1 solution to wash off the imprinted molecules to obtain molecularly imprinted polymer microspheres with selective recognition of caffeine artificial antibody.

[0062] The first step is the oxidation of horseradish peroxidase: Weigh 5 mg of horseradish peroxidase and dissolve it in 2 mL of freshly prepared acetate buffer containing sodium periodate, react in the dark for 30 minutes at room temperature, and add an appropriate amount of glycerol After c...

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Abstract

The invention relates to a preparation method for an artificial antibody for caffeine detection. Imprinted molecules located in an imprinted sphere layer are eluted with a prepared artificial antibody, so that a hole structure of which the structure, size and functional group are complementary to the structure, size and functional group of the imprinted molecules is formed in the imprinted spherelayer; the artificial antibody has specific recognition sites for enzyme-labeled antigen molecules, so that the artificial antibody can selectively recognize the enzyme-labeled antigen molecules; anda chemical immunoluminescence method is used to perform trace detection on the enzyme-labeled antigen molecules with a detection limit being 5.2*10<-11> mol.L<-1>. The preparation process of the artificial antibody includes the following four steps that: a horseradish peroxidase is oxidized; a horseradish peroxidase-labeled antigen is prepared, a caffeine antigen of a certain concentration gradient is configured, and the oxidized horseradish peroxidase is added dropwise, an enzyme-labeled antigen having the largest luminescence value is selected; a target molecular imprinted polymer is prepared with imprinted molecules, functional monomer, a cross-linking agent, and an initiator; and the target molecules are eluted, so that an artificial antibody capable of selectively recognizing caffeinecan be obtained.

Description

technical field [0001] The invention relates to the field of material science, in particular to a method for preparing an artificial antibody for caffeine chemiluminescence immunodetection. Background technique [0002] Caffeine is an alkaloid found in nature in the coffee, tea, cocoa and kola nuts and is a key ingredient in coffee, tea and so-called energy drinks. Caffeine has the effect of stimulating the central nervous system and keeps people excited. Appropriate use can expand coronary arteries, relax muscles, and relieve fatigue. Therefore, it is favored by modern stressed young people. However, large doses or long-term use can also cause damage to the human body, such as affecting sleep, causing ischemic heart disease, and even causing paroxysmal convulsions and bone tremors, damaging liver, stomach, kidney and other important internal organs, especially its It is addictive, and once the drug is stopped, a state of depression will appear. Therefore, it is urgent to e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/544
Inventor 高大明陈倩云吴梦瑶
Owner WUXI ZODOLABS BIOTECH
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