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Receptor reagent for homogeneous chemiluminiscence detection and application of receptor reagent

A homogeneous chemiluminescence, chemiluminescence technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, biological testing, etc., can solve problems such as extremely high sensitivity requirements

Pending Publication Date: 2020-12-22
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the progress of the detection industry, there is an increasing demand for ultrasensitive reagents, which not only require extremely high sensitivity, but also require a very wide detection range. It is difficult for the existing homogeneous chemiluminescence detection methods to meet the above detection requirements. condition

Method used

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  • Receptor reagent for homogeneous chemiluminiscence detection and application of receptor reagent
  • Receptor reagent for homogeneous chemiluminiscence detection and application of receptor reagent
  • Receptor reagent for homogeneous chemiluminiscence detection and application of receptor reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0158] Example 1: Preparation of the receptor particle solution of conjugated antibody I (PCT antibody)

[0159] (1) Preparation of antibody-coupled receptor particles with an average particle size of about 250 nm

[0160] 1.1 Preparation and characterization process of aldehyde-based polystyrene latex microspheres

[0161] 1) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min;

[0162] 2) Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride, dissolve them in 40ml of water to form an aqueous solution. This aqueous solution is added in the reaction system of step 1, continues to pass through N 2 30min;

[0163] 3) The temperature of the reaction system was raised to 70° C., and the reaction was carried out for 15 hours;

[0164] 4) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water by centrif...

Embodiment 2

[0192] Embodiment 2: the determination of kit sensitivity and upper limit of detection

[0193] The sensitivity point is defined as when the signal of concentration Cx is higher than the signal of twice the concentration of C0, that is, RLU(Cx)>2RLU(C0), then the sensitivity of the corresponding detection reagent is Cx. Define the detection limit point as the upper limit of the range determined using the method in the National Committee for Clinical Laboratory Standardization (NCCLS) Evaluation Protocol (EP) Series 6 document.

[0194] (1) Dilute the PCT antigen to 20pg / ml, 30pg / ml, 40pg / ml, 50pg / ml, 60pg / ml, 80pg / ml, 160pg / ml, 500pg / ml, 1000pg / ml, 5000pg / ml, 20000pg / ml ml, 50000pg / ml, 100000pg / ml and 200000pg / ml series concentration, the acceptor reagent ( The concentration is 100ug / ml), and then with the same biotin-labeled PCT monoclonal antibody 2 (diluted to 2ug / ml) and universal solution (reagent containing donor particles) to detect the above concentration series PCT a...

Embodiment 3

[0205] Example 3: Preparation of a series of acceptor particle solutions of conjugated antibody I (PCT antibody) with an average particle size of about 250 nm and different coefficients of variation in particle size distribution

[0206]According to the method described in (1) of Example 1, the receptor particle solution of conjugated antibody I with different coefficients of variation in particle size distribution was obtained, specifically:

[0207] Receptor particle 1: Gaussian distribution average particle size is 251.2nm, particle size distribution coefficient of variation C.V value=3.7%; Nicomp distribution is unimodal.

[0208] Receptor particle 2: the average particle size of Gaussian distribution is 254.9nm, the coefficient of variation C.V of particle size distribution=5.0%; the Nicomp distribution is unimodal.

[0209] Receptor particle 3: the average particle size of Gaussian distribution is 251.3nm, the coefficient of variation C.V of particle size distribution is...

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Abstract

The invention relates to the technical field of chemiluminiscence detection, and particularly discloses a receptor reagent for homogeneous chemiluminiscence detection and application of the receptor reagent. The receptor reagent comprises receptor particles capable of reacting with active oxygen to generate a detectable chemiluminiscence signal, wherein the particle size distribution variable coefficient C.V value of the receptor particles in the receptor reagent is greater than or equal to 5%. Compared with the prior art, the performance of a kit of the receptor reagent is greatly improved, and the kit not only has ultrahigh sensitivity, but also has a very wide detection range.

Description

technical field [0001] The invention belongs to the field of chemiluminescence detection, and in particular relates to an acceptor reagent for homogeneous chemiluminescence detection and its application. Background technique [0002] After more than half a century of development, immunoassays have developed many types. According to whether the substance to be tested is separated from the reaction system during the determination process, it can be divided into heterogeneous (Heterogenous) immunoassay and homogeneous (Homogeneous) immunoassay. Heterogeneous immunoassay refers to the operation process of introducing probes for labeling. Various related reagents need to be separated after mixing and reacting. The analyte is separated from the reaction system and then detected. It is the mainstream method in immunoassay. . Such as the well-known enzyme-linked immunosorbent assay (ELISA) and magnetic particle chemiluminescence. Homogeneous immunoassay refers to the direct measu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/58G01N33/532G01N21/76
CPCG01N21/76G01N33/532G01N33/54346G01N33/583G01N33/543G01N33/58
Inventor 康蔡俊吴晨杨阳刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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