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Kit for quantitatively detecting UCHL-1, and application thereof

A technology for UCHL-1 and detection reagents, which is applied in the field of kits for quantitative detection of UCHL-1, can solve the problems that the detection accuracy and repeatability cannot be improved, the upper limit of detection of the kit, the reaction speed and the detection sensitivity do not meet the requirements, etc.

Active Publication Date: 2021-04-09
SOPHONIX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection limit, reaction speed and detection sensitivity of the kit provided by the application do not meet the requirements, and the accuracy and repeatability of the detection cannot be improved, and the detection limit and linear relationship are not involved.

Method used

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  • Kit for quantitatively detecting UCHL-1, and application thereof
  • Kit for quantitatively detecting UCHL-1, and application thereof
  • Kit for quantitatively detecting UCHL-1, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] The preparation method of the enzyme-labeled UCHL-1 antibody conjugate and the UCHL-1 antibody magnetic particle conjugate comprises the following steps:

[0065] S1. Preparation of enzyme-labeled antibody conjugates

[0066] S1.1. Activation of antibodies:

[0067] S1.1.1 Activation step: Weigh 4-8mg of 2-iminosulfane hydrochloride (2-IT), dissolve it in buffer 1 to 13.76mg / mL, and set the molar ratio of 2-IT to antibody at 15-30:1 Add the 2IT solution to the antibody solution for activation (i.e. add 1mg antibody to 10-20μL of 2IT solution), shake and mix well, and react at room temperature for 30 minutes to obtain the activated antibody pre-product;

[0068] S1.1.2 Termination of activation step: add buffer 2 to the activated antibody pre-product solution at the ratio of 1 mg antibody to 5-20 μL buffer 2, react at room temperature for 10 min, use PD10 desalting column to remove excess 2IT, collect activated antibody, The activated antibody is obtained, and the acti...

Embodiment 2

[0084] Basic Example 2 A method for detection using a kit

[0085] Described detection method is:

[0086] (1) Immunological reaction: add 30uL sample, 50uL reagent B, and 50uL reagent A to the reaction well in sequence, and react at 37°C for 20 minutes;

[0087] (2) Magnetic separation and cleaning: add 300 μL of cleaning solution to the cleaning hole, magnetically suck the mixture containing magnetic particles out of the reaction hole, and demagnetize the cleaning hole M1; Bit M3 is subjected to magnetic separation and cleaning once respectively;

[0088] (3) Reading value: Add 150uL luminescent substrate to the reading well, and magnetically suck the mixture containing magnetic particles out of the cleaning hole M3, demagnetize the reading well, and the luminescent substrate catalyzed by alkaline phosphatase emits light. Use the instrument to detect the relative luminous intensity (RLU);

[0089] (4) A standard curve of UCHL-1 concentration-luminescence value can be obta...

Embodiment 3

[0116] Example 3 A kit for quantitative detection of UCHL-1

[0117]

[0118]

[0119] The detection reagent strip is composed of reagent A, reagent B, cleaning solution, luminescent substrate, measuring and reading well, elution sleeve, and tip;

[0120] The reagent A is a solution prepared by using the enzyme-labeled UCHL-1 antibody conjugate as a raw material and thoroughly mixing it with buffer B;

[0121] Buffer B component Weighing amount tris hydroxymethyl amino methane 15.0g Sodium chloride 9.0g bovine serum albumin 30g glycerin 5.0g Glycine 40g pH value 8.2 purified water Dilute to 1000mL

[0122] The reagent B is a solution prepared by using the UCHL-1 antibody magnetic particle conjugate as a raw material and thoroughly mixing it with buffer C.

[0123] Buffer C component Weighing amount Disodium hydrogen phosphate dodecahydrate 5.8g Sodium dihydrogen phosphate 0.57g Sodiu...

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Abstract

The invention provides a kit for quantitatively detecting UCHL-1, and application thereof, and belongs to the technical field of biological detection. The kit is composed of a detection reagent strip, a calibration product, a quality control product and a two-dimensional code; the detection reagent strip comprises a reagent A and a reagent B; the reagent A is a solution prepared by taking an enzyme-labeled UCHL-1 antibody conjugate as a raw material and fully and uniformly mixing the enzyme-labeled UCHL-1 antibody conjugate with a buffer solution B; and the reagent B is a solution prepared by taking a UCHL-1 antibody magnetic particle conjugate as a raw material and fully and uniformly mixing the UCHL-1 antibody magnetic particle conjugate by using a buffer solution C. In the implementation process, the specific components of the buffer solution B and the buffer solution C and the mass ratio of the components are controlled, so the research on indexes such as appearance, accuracy, blank limit, detection limit, quantification limit, linearity, repeatability, difference between calibration product and quality control product bottles and the like of the prepared kit meets the requirements, the kit is verified to be qualified on a full-automatic chemiluminescence immunoassay analyzer, and the measurement result meets the enterprise standard requirements.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a kit for quantitative detection of UCHL-1 and an application thereof. Background technique [0002] Protein gene product 9.5 (protein gene product 9.5, PGP9.5), also known as ubiquitin carboxyl-terminal hydrolase (UCH-L1), is a class of cysteine ​​hydrolase composed of 223 amino acids. The molecular weight is about 24800Da. These enzymes are involved in the ubiquitination and deubiquitination of proteins in cells through the ATP-dependent proteasome pathway. Mutations and deletions of the UCH-L1 gene are closely related to familial Parkinson's disease and axonal dystrophy. Traumatic brain injury (TBI) is an injury to the brain caused by an external force that disrupts normal brain function, resulting in impairment of a person's cognitive abilities or physical functions. Among all types of TBI, the most common sequelae are headache (47.9%) and memory a...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/573G01N2800/28
Inventor 刘聪冯玉静李博飞郑兴华
Owner SOPHONIX CO LTD
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