A kind of preparation method of magnetic particle chemiluminescence reagent

A magnetic particle and light-proof technology, which is applied in the field of immunological technology detection, can solve the problem that the linear range of the reagent is not wide enough

Active Publication Date: 2018-07-10
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the linear range of the reagent is not wide enough, and high-value samples need to be diluted in advance

Method used

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  • A kind of preparation method of magnetic particle chemiluminescence reagent
  • A kind of preparation method of magnetic particle chemiluminescence reagent
  • A kind of preparation method of magnetic particle chemiluminescence reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: control method

[0041] 1. Preparation of magnetic particles coated with myoglobin monoclonal antibody

[0042] 1.1 Thoroughly mix magnetic particles (Merck, particle size 0.8 to 1.2 μm, product number 39433087), take 10 mg into a test tube, magnetically separate for 1 minute, and remove the supernatant. Add 1ml of binding buffer (0.1M MES, pH 5.0), mix well, then magnetically separate for 1 minute, then remove the supernatant, add 1ml of binding buffer, and mix the magnetic particles evenly.

[0043] 1.2 Add 100 μg myoglobin monoclonal antibody and mix for 30 minutes at room temperature (22°C to 28°C).

[0044] 1.3 Add 100 μl of 10 mg / ml EDC (prepared immediately before use, the solvent is ice binding buffer) and mix well at room temperature for 3 hours in the dark. Remove the supernatant by magnetic separation, add washing buffer (50mM TBS, pH 7.4), mix and remove the supernatant by magnetic separation, repeat three times.

[0045] 1.4 Add 1ml of quen...

Embodiment 2

[0049] Example 2: Optimized magnetic particle coating method

[0050] 1. Preparation of magnetic particles coated with myoglobin monoclonal antibody

[0051] 1.1 Thoroughly mix magnetic particles (Merck, particle size 0.8 to 1.2 μm, product number 39433087), take 10 mg into a test tube, magnetically separate for 1 minute, and remove the supernatant. Add 1ml of binding buffer (0.1M MES, pH 5.0), mix well, then magnetically separate for 1 minute, then remove the supernatant, add 1ml of binding buffer, and mix the magnetic particles evenly.

[0052] 1.2 Add 100 μl of 10 mg / ml EDC (ready to use, the solvent is ice binding buffer), mix at room temperature (22°C to 28°C) for 30 minutes.

[0053] 1.3 Add 100 μg myoglobin monoclonal antibody, and mix well at room temperature for 3 hours in the dark. Remove the supernatant by magnetic separation, add washing buffer (50mM TBS, pH 7.4), mix and remove the supernatant by magnetic separation, repeat three times.

[0054] 1.4 Add Biolipi...

Embodiment 3

[0061] Example 3. Encapsulation of Magnetic Particles

[0062] In order to compare the blocking effect of the blocking solution, after step 1.3 of Example 2, the coated magnetic particles were washed with washing buffer and divided into two, and 200 μl of Biolipidure-502 or Biolipidure-1002 were added for blocking.

[0063] The results (Table 1) show that the signal-to-noise ratio of the magnetic particles blocked by Biolipidure-1002 in the reaction is 5.304; the sensitivity is better, and the high-value linear ratio is also better. Because Biolipidure contains polymers with different molecular weights, the sealing effect of magnetic particles is better.

[0064] The magnetic particles blocked by Biolipidure-1002 were placed at 37°C for 15min, 30min and 60min respectively. The test results (Table 2) show that the performance of the magnetic particles treated at 37°C for 15 minutes is not significantly different from that of the untreated ones in the reaction.

[0065] Howeve...

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Abstract

The invention relates to a preparation method of a magnetic particle chemiluminescence reagent, and also relates to a detection kit of human myoglobins. The method comprises the following steps: mixing magnetic particles with a cross-linking agent, and combining the obtained mixture with an antibody to be coated. A polymer is adopted by the method in the invention to close the magnetic particles. The detection regent prepared through the method combines respective advantages of chemiluminescence and a magnetic particle technology, so the magnetic particles make the kit have high specificity and sensitivity, large detection range and fast detection time.

Description

technical field [0001] The disclosure belongs to the field of immune technology detection, and in particular relates to a preparation method of a magnetic particle chemiluminescence kit and the kit prepared by the method; the disclosure also relates to a quantitative kit for detecting myoglobin in human serum. Background technique [0002] Myoglobin is a protein present in the cytoplasm of cardiac and skeletal muscle with a molecular weight of 17.8kD. [0003] After muscle cell damage, myoglobin is rapidly released into the circulation. Determination of myoglobin in serum is one of the important indicators for the diagnosis of acute myocardial infarction (AMI), reinfarction and successful reperfusion after thrombolytic therapy. Concentrations Myoglobin concentrations rise about two hours after the onset of symptoms, and thus serve as an early indicator for the diagnosis of myocardial infarction. Depending on the reperfusion therapy, after the onset of infarction, circulati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/543G01N33/68
CPCG01N33/531G01N33/54326G01N33/6893G01N2333/805G01N2800/324G01N2800/50
Inventor 闫欣孙国敬崔伟刘希
Owner BEIJING STRONG BIOTECH INC
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