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Composition for antigen detection and preparation method

A technology of antigen detection and composition, which is applied in the field of immune detection and can solve problems such as abnormal detection values

Inactive Publication Date: 2021-05-07
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, commonly used markers in chemiluminescent immunoassays include acridinium esters, alkaline phosphatase, horseradish peroxidase, luminol, and tripyrimidine ruthenium, among which alkaline phosphatase as a marker appeared earlier and is relatively Mature technology platform, complete raw material process system, mature process system and relatively easy-to-implement equipment, but alkaline phosphatase has shown different effects in different antigen / antibody detection, which is obviously affected by the antigen / antibody itself. The impact of molecular weight, for antigens with small molecular weight, the use of alkaline phosphatase has obvious defects, which will directly lead to abnormal detection values, so solving the problems of alkaline phosphatase in labeling is important for the improvement and improvement of chemiluminescence methods. The further advancement of clinical research is of great significance

Method used

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  • Composition for antigen detection and preparation method
  • Composition for antigen detection and preparation method
  • Composition for antigen detection and preparation method

Examples

Experimental program
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Effect test

preparation example Construction

[0066] 2. Preparation of biotinylated anti-reagent

[0067] A: Preparation of biotinylation reagent

[0068] In 100mM carbonate solution, cTnI antibody 1 was desalted and concentrated to a final concentration of 2mg / ml; biotin was dissolved in DMSO, and biotin was added to the first cardiac troponin I antibody at a ratio of 10:1 at 37°C Coupling for 2 hours, using a desalting column to remove uncoupled biotin to obtain biotin resistance reagent;

[0069] B: Dilute biotinylated anti-reagent

[0070] Add anti-reagent diluent with the following components to the biotin anti-reagent: 50mM This-HCl buffer, pH=7.4; 0.9% NaCl, 0.5% BSA, 0.09% Proclin-300, 0.1% Tween-20, 1% glycerol, Make the final concentration 1 μg / mL.

[0071] 3. Preparation of magnetic particle composites

[0072] The carboxyl magnetic beads were activated with EDC and washed four times; the antibody was activated with TCEP, concentrated in an ultrafiltration tube and then replaced with PBS buffer; the carboxy...

Embodiment 1

[0084] The impact of the first activator of embodiment 1 on detection result

[0085] Reagent components: 3 mg / mL alkaline phosphatase, 0.66 mg / mL streptavidin, 30 mg / mL first activator, 6.6 mg / mL SDPD.

[0086] The first activator in reagent component 1 is SM (PEG) 4 ;

[0087] The first activator in reagent component 2 is SM (PEG) 8 ;

[0088] The first activator in reagent component 3 is SM (PEG) 12 ;

[0089] The first activator in reagent component 4 is SM (PEG) 24 ;

[0090] The first activator in reagent component 5 is SMCC;

[0091] No first activator was added to reagent component 6.

[0092] The specific operation in this example: the coupling ratio of streptavidin to alkaline phosphatase is 1:4.5, and the ratio of the first activator to the conjugate is 10:1.

[0093] Mix 3mg / mL of alkaline phosphatase with 30mg / mL of the first activator to activate alkaline phosphatase, mix 0.66mg / mL of streptavidin with 6.6mg / mL of SDPD to activate streptavidin , (the fi...

Embodiment 2

[0112] The influence of the second activator of embodiment 2 on detection result

[0113] Reagent components: 3mg / mL alkaline phosphatase, 0.66mg / mL streptavidin, 30mg / mL SM (PEG) 12 , 6.6 mg / mL of the second activating reagent.

[0114] The second activator in reagent component 7 is DTT;

[0115] The second activator in reagent component 8 is TCEP;

[0116] The second activator in reagent component 9 is 2-IT;

[0117] No second activator was added to reagent component 10.

[0118] In this embodiment, the specific operation is to mix 3mg / mL of alkaline phosphatase with 30mg / mL of SM(PEG)12 to activate alkaline phosphatase, and 0.66mg / mL of streptavidin and 6.6mg / mL of The two activators are mixed to activate the streptavidin, (the second activator is not added in the reagent component 10, the streptavidin is directly added after the alkaline phosphatase is activated) and then the activated streptavidin is Mix with activated alkaline phosphatase to prepare alkaline phosphata...

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Abstract

The invention relates to the field of immunodetection, in particular to a composition for antigen detection and a preparation method, and discloses the composition for antigen detection, which realizes high-sensitivity and high-accuracy detection of antigens by utilizing a long-chain activator to amplify the length of an antigen-antibody connecting arm.

Description

technical field [0001] The invention relates to the field of immunodetection, in particular to a composition and a preparation method for antigen detection. Background technique [0002] Chemiluminescent immunoassay is an analytical technique that combines chemiluminescent assay technology with immune reaction. Because it has both high sensitivity of chemiluminescent detection and high specificity of immune reaction, it is widely used in various antigens, semi- In the detection and analysis of antigens, antibodies, hormones, enzymes, fatty acids, vitamins and drugs, it is a newest immunoassay technology developed after enzyme-linked immunoassay, fluorescent immunoassay and time-resolved fluorescent immunoassay. [0003] At present, commonly used markers in chemiluminescent immunoassays include acridinium esters, alkaline phosphatase, horseradish peroxidase, luminol, and tripyrimidine ruthenium, among which alkaline phosphatase as a marker appeared earlier and is relatively ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/58G01N33/53
CPCG01N33/5306G01N33/54326G01N33/54393G01N33/581
Inventor 陈亮德郑旻亮
Owner ZYBIO INC
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