Antibody coating process and myohemoglobin determination reagent kit made with same

A myoglobin and antibody technology, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of high cost and complex preparation process of antibody microspheres, and achieve the effect of improving efficiency, remarkable effect and simple operation

Active Publication Date: 2016-12-07
SICHUAN XINCHENG BIOLOGICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is: the coupling efficiency of existing antibodies and microspheres is usually improved by bridging various chemical substances, which leads to more complicated preparation process and higher cost of antibody microspheres; the purpose of the present invention It is to provide an antibody coating process that can effectively improve the coupling efficiency between antibodies and microspheres by using only one activator, and to provide a myoglobin assay kit including antibody microspheres prepared by this process

Method used

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  • Antibody coating process and myohemoglobin determination reagent kit made with same
  • Antibody coating process and myohemoglobin determination reagent kit made with same
  • Antibody coating process and myohemoglobin determination reagent kit made with same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] An antibody coating process, the specific process is as follows:

[0031] Step 1, the microspheres and the activator are dissolved in the buffer solution of pH6.0 respectively; the buffer solution in this step adopts potassium dihydrogen phosphate-sodium hydroxide buffer solution, and the microspheres adopt polystyrene latex microspheres , the activator is EDAC.

[0032] Step 2, using an activator to activate the carboxyl groups on the surface of the microspheres; that is, adding the buffer solution containing EDAC to the dissolved microspheres to activate the hydroxyl groups on the surface of the microspheres, and the activation time is 30 minutes.

[0033] Step 3, then adding the antibody to the activated microspheres for coupling, the coupling time is 1h;

[0034] Step 4, adding potassium dihydrogen phosphate-sodium hydroxide buffer solution with pH 8.0 to the antibody-coupled microspheres to continue the reaction, the reaction time is 1 h;

[0035] Step 5. Add cas...

Embodiment 2

[0038] This embodiment is a comparative example of embodiment 1. The difference between this embodiment and embodiment 1 is that this embodiment has modified the specific operation process after step 3. At the same time, the pH value in the whole process of this embodiment is different. The specific process after step three is as follows:

[0039] Step 3, then add the antibody to the activated microspheres for coupling, and the coupling time is 2 hours;

[0040] Step 4: Add casein and glycine to complete the blocking after the reaction is completed, and the blocking time is 30 minutes; after the blocking is completed, add potassium dihydrogen phosphate-sodium hydroxide buffer solution with the same pH value as in step 1, and adjust the pH value to 7.5, Then add trehalose and xylitol as protective agents and store it.

[0041] In this embodiment, buffer solutions with different pH values ​​were used for experiments, and the values ​​of the calibration curve after coating with ...

Embodiment 3

[0046] The difference between this example and Example 1 and Example 2 is that the activators used in this example are selected as EDAC and Sulfo-NHS. Activation, other operation steps are exactly the same as embodiment 1 and embodiment 2.

[0047] This example also gives the values ​​of the calibration curve after coating by the two activation coating methods in Example 1 and Example 2. The values ​​are shown in Table 1, and the coupling under different pH conditions is detected. Efficiency graphs such as figure 2 shown.

[0048] Table 2

[0049]

[0050]

[0051] pass figure 2 The results show that the coupling efficiency between the antibody and the microspheres is improved to a certain extent by using the pH gradient coating method of the present invention relative to the fixed pH value. But by comparing the test data of embodiment 2 and embodiment 3, it can be seen that when only one activator is used, its coupling efficiency is significantly higher than when ...

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Abstract

The invention discloses an antibody coating process and a myohemoglobin determination reagent kit made with the same. The problem that in the prior art, bridging of various chemical substances is generally adopted to improve the antibody and microsphere coupling efficiency, and consequently an antibody microsphere preparation process is more complex and higher in cost is solved. The process includes the steps that firstly, microspheres and an activator are dissolved in a buffer solution with the pH of 6.0 respectively; secondly, carboxyl on the surfaces of the microspheres is activated with an activator to obtain activated microspheres; thirdly, antibodies are added into the activated microspheres for coupling; fourthly, a buffer solution with the pH of 8.0 is added into the activated microspheres coupled with the antibodies to continue a reaction; fifthly, after the reaction is completed, a sealing agent is added to complete sealing. The process has the advantages that the chemical coupling efficiency is greatly improved, the operation steps are simplified, and the production cost is reduced.

Description

technical field [0001] The invention relates to a coating process, in particular to an antibody coating process, and provides a myoglobin assay kit made by the process. Background technique [0002] Myoglobin is a small molecular protein, its molecular structure is similar to hemoglobin, and it has the function of transporting and storing oxygen in muscle cells. Human cardiac muscle and skeletal muscle contain a large amount of myoglobin, which is rarely found in the blood of normal people, and is mainly metabolized and excreted by the kidneys. When the myocardium or striated muscle is damaged, myoglobin is released into the blood, and the myoglobin in the serum can be significantly increased, so the determination of myoglobin can be used for the diagnosis of some myopathy and heart disease, such as acute myocardial infarction (AMI), acute muscle injury, acute and chronic renal failure, severe congestive heart failure, prolonged shock, muscular dystrophy, muscular atrophy a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54313G01N33/6887G01N2333/4712
Inventor 林婷婷林源谭韬
Owner SICHUAN XINCHENG BIOLOGICAL CO LTD
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