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Electrochemiluminescence kit for detecting Titin antibody and preparation method of kit

A titin and electrochemical technology, applied in the field of immunoassay, can solve the problems of low-risk and accurate quantitative detection of Titin antibody methods or kits, etc., and achieve the effects of rapid detection, high detection sensitivity, and wide linear range

Pending Publication Date: 2020-04-21
江苏三联生物工程股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is: in order to solve the technical problem that there is no rapid, low-risk, and accurate quantitative detection method or kit for Titin antibody in the prior art, an electrochemiluminescence kit for detecting Titin antibody and its preparation are provided. Law

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  • Electrochemiluminescence kit for detecting Titin antibody and preparation method of kit
  • Electrochemiluminescence kit for detecting Titin antibody and preparation method of kit
  • Electrochemiluminescence kit for detecting Titin antibody and preparation method of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1 detects the preparation method of the electrochemiluminescence kit of Titin antibody

[0023] (1) Preparation of streptavidin-coupled magnetic particle working solution

[0024] The streptavidin-coupled magnetic particle suspension was magnetically separated to remove the supernatant, and resuspended to a concentration of 0.5 mg / mL with a pH of 6.8 and a concentration of 0.1 mol / L PBS buffer containing 0.5 wt% bovine serum albumin, 0.05 wt% Triton X-100, 0.05 wt% proclin300 and 0.05 wt% sodium azide.

[0025] (2) Preparation of biotin-labeled Titin antigen

[0026] Take 1 mg of recombinant Titin antigen, add an appropriate amount of PBS buffer (0.05M, pH7.0-7.6) to adjust the total antibody concentration to 1 mg / ml, and put it into a dialysis bag for dialysis, changing the dialysate every 3-4 hours, Replace 3-4 times, transfer the antigen to a centrifuge tube or cryopreservation tube after dialysis;

[0027] Accurately weigh 1 mg of biotin activated by N-h...

Embodiment 2

[0041] Example 2 The method of using the kit of the present invention to detect Titin antibody uses a fully automatic electrochemiluminescence immunoassay analyzer as a detection instrument, and the kit is loaded on the instrument for detection, and the steps are as follows:

[0042] Add the sample (or calibrator or quality control), biotin-labeled Titin antigen working solution and ruthenium-tripyridyl-labeled anti-human IgG antibody working solution into the cuvette, and incubate at 37°C for 10 minutes to form antigen-antibody-antibody Antibody complex solution; add streptavidin-coated magnetic particle working solution to the antigen-antibody-antibody complex solution, and incubate at 37°C for 10 minutes to form a magnetic complex suspension;

[0043] The magnetic composite suspension is placed in a magnetic field, and the cleaning solution flows through to wash the magnetic composite;

[0044] The electrochemiluminescence substrate solution is successively injected into th...

Embodiment 3

[0046] The research of embodiment 3 reagent sensitivity

[0047] Reagent sensitivity is determined according to the lowest detection limit (LOB), which is carried out according to the following experimental method. Detect the zero-concentration calibrator 20 times, get the signal value (RLU) of 20 measurement results, calculate the mean M and standard deviation SD, and obtain the RLU value corresponding to M+2SD, according to the zero-concentration calibrator and the adjacent concentration The concentration-RLU value results between the calibrator (adjacent concentration is 80pg / mL) are subjected to two-point regression fitting to obtain a linear equation, and the RLU value corresponding to M+2SD is brought into the above equation to calculate the corresponding Concentration, the lower limit of detection (LOB).

[0048] The measurement results in Table 1 below show that the detection limit (LOB) of the reagent detected by the above method is 41.5 pg / mL, and according to this ...

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Abstract

The invention relates to an electrochemiluminescence kit for detecting a Titin antibody in blood and a preparation method of the electrochemiluminescence kit. The prepared kit comprises a streptavidincoupled magnetic particle working solution, a biotin labeled Titin antigen working solution, a ruthenium terpyridyl labeled anti-human IgG antibody working solution, a working solution of a calibrator and / or a quality control product of the Titin antibody, a tripropylamine-containing electrochemiluminescence substrate solution and a cleaning solution. The kit adopts electrochemiluminescence, a streptavidin-biotin signal amplification system is utilized, the detection sensitivity is high, the linear range is wide, the detection result repeatability is high, and accurate quantification of the Titin antibody can be realized.

Description

technical field [0001] The invention relates to an electrochemiluminescence kit and a preparation method for detecting titin antibody (Titin antibody), which belong to the technical field of immune analysis. Background technique [0002] Autoimmune diseases are diseases in which the immune system reacts to the components of its own body, causing damage. Under normal circumstances, the immune system only responds to foreign substances that invade the body, such as bacteria, viruses, parasites, and transplants, and destroys or rejects these foreign substances. Under the influence of certain factors, some abnormalities appear in the tissue components of the body or the immune system itself, causing the immune system to mistake its own components as foreign objects to attack. At this time, the immune system will produce antibodies against some of the body's own components and active lymphocytes, which will damage and destroy its own tissues and organs, leading to disease. If l...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N27/26G01N21/76
CPCG01N21/76G01N27/26G01N33/54326G01N2800/24
Inventor 孟君泮锋纲丁俊杰施启尧
Owner 江苏三联生物工程股份有限公司
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