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A kit for detecting tumor m2-type pyruvate kinase and its preparation method

A technology for pyruvate kinase and tumors, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of easy deviation of test results, many influencing factors, and many detection steps, and achieve objective, sensitive and broad test results Application prospect, effect of long fluorescence lifetime

Active Publication Date: 2020-03-10
广州春康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the tumor M2-PK kits developed at home and abroad all use the enzyme immunoassay method. This method has many detection steps and many influencing factors in the operation process. The detection results are easy to cause deviations, and it is time-consuming and laborious.
Although quantum dot biological labeling has been reported, quantum dot-conjugated antibodies and purification have disadvantages such as complicated operation, low recovery rate, and difficulty in batch detection and scale-up.

Method used

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  • A kit for detecting tumor m2-type pyruvate kinase and its preparation method
  • A kit for detecting tumor m2-type pyruvate kinase and its preparation method
  • A kit for detecting tumor m2-type pyruvate kinase and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] This embodiment provides a method for preparing a quantum dot-labeled capture antibody, which includes the following steps:

[0051] S1) Activation of quantum dots: using N-hydroxysulfosuccinimide to activate the carboxyl groups on the surface of CdSe / ZnS quantum dots;

[0052] S2) modifying the capture antibody with monophosphate sugar to obtain an activated capture antibody;

[0053] S3) Equilibrate the water-soluble CdSe / ZnS (1-10 μM) quantum dots with a concentration of carboxyl group to room temperature, add EDC solution (9.38%), N-hydroxyl sulfosuccinimide solution (10%) and BSA solution (20-200mg / mL), shake well, add activated capture antibody (10-100μg / ml), shake well and incubate for 30-45min, add EDC solution (9.38%), N-hydroxythio Alternative succinimide solution (10%); after incubation, add methanol, mix and avoid light and shake for 1.5-2 hours; then add β-mercaptoethanol to terminate, and then add β-mercaptoethanol to stabilize the quantum dots after term...

Embodiment 2

[0056] This example provides a method for preparing a quantum dot-labeled capture antibody, which differs from Example 1 in that: in step S3, when the activated quantum dot is coupled to the activated capture antibody, the pH of the solution is 10, and the activated The mass ratio of quantum dots to twice added EDC is 7, the mass ratio of activated quantum dots to twice added N-hydroxysulfosuccinimide is 8; the mass ratio of activated quantum dots to activated capture antibodies It is 1:7.

Embodiment 3

[0058] This example provides a method for preparing a quantum dot-labeled capture antibody, which differs from Example 1 in that: in step S3, when the activated quantum dot is coupled to the activated capture antibody, the pH of the solution is 11, and the activated The mass ratio of quantum dots to EDC added twice is 10, the mass ratio of activated quantum dots to N-hydroxysulfosuccinimide added twice is 10; the mass ratio of activated quantum dots to activated capture antibody It is 1:10.

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Abstract

The invention discloses a kit for inspecting tumour M2 type pyruvate kinase and a preparation method of the kit. The kit comprises a test paper card. The test paper card comprises a bottom plate, thebottom plate is provided with a sample pad, a combination pad, a nitrocellulose membrane and an absorbent pad which are sequentially connected at the ends in the sample flowing direction, capture antibodies marked by quantum dots are adsorbed by the combination pad, the nitrocellulose membrane is successively provided with an inspection belt and a quality control belt in the sample flowing direction, the inspection belt is covered with inspection antibodies, and the quality control belt is covered with IgG; the capture antibodies and the inspection antibodies are anti-tumour M2 type pyruvate kinase antibodies, and the capture antibodies and the inspection antibodies are combined with different epitopes of the tumour M2 type pyruvate kinase; and before the capture antibodies are marked by the quantum dots, the capture antibodies are modified with monophosphate sugar. The kit has the advantages that high sensitivity and specificity are achieved, operation is fast, simple and convenient,the result is accurate, and affordability is achieved.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis, and in particular relates to a kit and a preparation method for detecting tumor M2 type pyruvate kinase. Background technique [0002] Colorectal cancer is one of the three most common malignant tumors in the world, and its mortality rate ranks third. In recent years, with the improvement of people's living standards and the change of diet structure, the incidence rate has shown an obvious upward trend. However, the early diagnosis of colorectal cancer at home and abroad is still at a low level, and about half of the patients have entered the middle and late stages of colorectal cancer when they are clearly diagnosed. Therefore, improving the early diagnosis of colorectal cancer is a key issue that urgently needs to be solved at home and abroad. At present, there are several commonly used techniques for early diagnosis of colorectal cancer: (1) Fecal occult blood test is a commonly u...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/573G01N33/533
Inventor 何凤屏刘布
Owner 广州春康生物科技有限公司
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