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Thymidine kinase 1 magnetic particle chemiluminiscence assay kit and preparation method thereof

A thymidine kinase and chemiluminescence technology, applied in measurement devices, scientific instruments, instruments, etc., can solve the problems of labor and time consumption, low sensitivity and linear range, unable to meet clinical needs, etc., to reduce errors, specificity and accuracy. Sensitivity improvement, the effect of contributing to detection sensitivity

Pending Publication Date: 2020-04-21
SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method usually relies on manual operation, which is labor-intensive and time-consuming. At the same time, its detection sensitivity and linear range are also low. It is mainly used for laboratory analysis. It has certain limitations in clinical detection applications and is far from meeting clinical needs.
At present, there is still a lack of TK1 assay reagents with fast speed, good stability, high sensitivity and strong specificity in the market, especially automated detection reagents

Method used

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  • Thymidine kinase 1 magnetic particle chemiluminiscence assay kit and preparation method thereof
  • Thymidine kinase 1 magnetic particle chemiluminiscence assay kit and preparation method thereof
  • Thymidine kinase 1 magnetic particle chemiluminiscence assay kit and preparation method thereof

Examples

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Effect test

Embodiment 1

[0059] Rabbit anti-TK1 polyclonal antibody coated p-toluenesulfonyl magnetic particles

[0060] The magnetic particles coated with rabbit anti-TK1 polyclonal antibody are formed by linking p-toluenesulfonyl magnetic particles with amino groups of rabbit anti-TK1 polyclonal antibody. The specific synthesis steps of the complex are as follows:

[0061] (1) Weigh 3.1g of boric acid, dissolve it in 1L of deionized water, adjust the pH to 9.6, and make buffer solution A;

[0062] (2) Weigh 7.9g Tris-HCl, dissolve it in 1L deionized water, adjust the pH to 7.6, and make buffer solution B;

[0063] (3) Take 10g of bovine serum albumin (BSA), dissolve it in 1L of the above-mentioned buffer solution B, and make buffer solution C;

[0064] (4) Add 1g of p-toluenesulfonyl magnetic particles and 10mg of rabbit anti-TK1 polyclonal antibody to 100ml of buffer solution A, and then react the mixed solution at room temperature (20-25°C) for 16-24 hours. During the reaction, the container Rol...

Embodiment 2

[0069] Mouse anti-TK1 monoclonal antibody coated carboxylated magnetic particles

[0070] The magnetic particles coated with mouse anti-TK1 monoclonal antibody are formed by linking carboxyl magnetic particles with amino groups of mouse anti-TK1 monoclonal antibody. The specific synthesis steps of the complex are as follows:

[0071] (1) Weigh 9.7g of 2-(N-morpholine)ethanesulfonic acid, dissolve it in 1L of deionized water, adjust the pH to 5.5, and make buffer solution A;

[0072] (2) Weigh 7.9g Tris-HCl, dissolve it in 1L deionized water, adjust the pH to 7.6, and make buffer solution B;

[0073] (3) Take 10g of bovine serum albumin (BSA), dissolve it in 1L of the above-mentioned buffer solution B, and make buffer solution C;

[0074] (4) Add 1 g of carboxyl magnetic particles and 10 mg of mouse anti-TK1 monoclonal antibody to 100 ml of buffer solution A, then add 0.5 g of EDC·HCl, and then react the mixed solution at room temperature (20-25° C.) for 2 hours. Place the co...

Embodiment 3

[0079] Preparation of Alkaline Phosphatase-labeled Goat Anti-TK1 Polyclonal Antibody Enzyme Conjugate

[0080] Alkaline phosphatase-labeled goat anti-TK1 polyclonal antibody enzyme conjugate is formed by linking alkaline phosphatase and TK1 antibody. The specific synthesis steps of the complex are as follows:

[0081] (1) Weigh 2.9g of sodium bicarbonate and 1.5g of sodium carbonate, dissolve them in 1L of deionized water, adjust the pH to 9.5, and make buffer solution A;

[0082] (2) Dissolve 100mg of alkaline phosphatase in 5ml of deionized water and mix well, then add 100mg of sodium periodate into it, shake and mix until completely dissolved, and activate the reaction at 2-8°C for 30min;

[0083](3) Add 0.36ml of ethylene glycol to the alkaline phosphatase activation solution in step (2), mix well and place it at 2-8°C for 2 hours to terminate the activation reaction;

[0084] (4) Add 100mg of goat anti-TK1 polyclonal antibody to the alkaline phosphatase reaction solution...

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Abstract

The invention relates to a thymidine kinase 1 magnetic particle chemiluminiscence assay kit and a preparation method thereof. The kit comprises a reagent A, a reagent B and a thymidine kinase 1 (TK1)calibrator, the reagent A is a magnetic particle suspension coated with an anti-thymidine kinase 1 (TK1) antibody, and the reagent B is an alkaline phosphatase labeled anti-thymidine kinase 1 (TK1) antibody enzyme conjugate reagent. Compared with the prior art, the invention provides the method for determining the content of thymidine kinase 1 (TK1), and by utilizing the kit provided by the invention, the content of thymidine kinase 1 (TK1) in a sample can be accurately, sensitively and quickly determined according to the principle of a double-antibody sandwich method.

Description

technical field [0001] The invention relates to a biological reagent, in particular to a detection kit, in particular to a kit for measuring human serum thymidine kinase 1 content with high sensitivity and strong specificity and a preparation method thereof. Background technique [0002] Thymidine kinase 1 (Thymidine Kinase 1, TK1) Thymidine kinase (TK1) is one of the key enzymes in DNA synthesis, under the conditions of participation of magnesium ions and ATP, it can catalyze the conversion of thymidine (TdR) into deoxygenation Thymidine monophosphate (TMP), and then formed thymidine triphosphate (TTP, one of the four necessary deoxynucleotides in DNA synthesis) to participate in DNA synthesis. The increase of TK1 level is positively correlated with the synthesis of DNA. In the cell cycle, TK1 begins to increase in the late G1 phase, reaches the highest level in the S phase, and begins to decrease in the G2 phase. Due to the special correlation between TK1 and the S phase o...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/543
CPCG01N33/573G01N33/54326G01N2333/916Y02P20/55
Inventor 罗紫臣
Owner SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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