Preparation method of serum carbon dioxide determination reagent ball and reagent ball

A technology of carbon dioxide and reagents, which is applied in the preparation method of reagent balls for serum carbon dioxide determination and the field of reagent balls, can solve the problems of difficulty in applying instant diagnosis of patients, short validity period of liquid reagents, and use restrictions, and achieves guaranteed shape and reconstitution solubility, Good shape and effect of reconstitution solubility, high stability and precision

Pending Publication Date: 2021-08-20
GENRUI BIOTECH INC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the enzymatic method used clinically to measure serum carbon dioxide, on the one hand, the reagent solution needs to add a circulatory regeneration system of reducing coenzyme I (NADH), and the stability is poor
On the other hand, the stability of the dry powder reagent after reconstitution is poor, the validity period of the liquid reagent is short, and it relies on a large-scale automatic biochemical analyzer, which is difficult to apply to the patient's immediate diagnosis (point-of-care, POCT). certain limit
Although there are existing freeze-dried reagent balls used to detect other items, the shape of the existing freeze-dried reagent balls is poor, and it is difficult to completely freeze-dry, resulting in lower stability and accuracy.

Method used

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  • Preparation method of serum carbon dioxide determination reagent ball and reagent ball
  • Preparation method of serum carbon dioxide determination reagent ball and reagent ball
  • Preparation method of serum carbon dioxide determination reagent ball and reagent ball

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preparation example Construction

[0033] The embodiment of the present invention provides a method for preparing a serum carbon dioxide measurement reagent ball, which can prepare a spherical serum carbon dioxide measurement reagent. Please refer to figure 1 , the method includes the following steps:

[0034] S10: Mix buffer, phosphoenolpyruvate, reduced coenzyme I, malate dehydrogenase, phosphoenolpyruvate carboxylase, magnesium salt, stabilizer, excipient and water to form a mixed solution, And adjust the pH value of the mixed solution to a preset value.

[0035] In this example, buffer, phosphoenolpyruvate, reduced coenzyme I, malate dehydrogenase, phosphoenolpyruvate carboxylase, magnesium salt, stabilizer, excipient and water (each component), and then thoroughly mix each component to obtain a mixed solution. The pH value of the mixed solution can be tested by methods such as pH test paper, and the pH value of the mixed solution can be adjusted to a preset value by adding sodium hydroxide. Wherein, the...

Embodiment 1

[0063] In this embodiment, the buffer includes Tris buffer, the stabilizer includes bovine serum albumin, the excipient includes mannitol, and the magnesium salt includes magnesium sulfate. The preparation method of serum carbon dioxide measurement reagent ball is as follows: a certain amount of Tris buffer, bovine serum albumin, mannitol, magnesium sulfate, phosphoenolpyruvate, reduced coenzyme Ⅰ, malate dehydrogenase, phosphoenol Formula pyruvate carboxylase and water are mixed to form a mixed solution, and the droplets of the mixed solution are dropped into liquid nitrogen to form ice balls with a volume of about 3.0ul, and the ice balls are freeze-dried to prepare reagent balls for measuring carbon dioxide in serum. And the microfluidic chip is prepared by using the serum carbon dioxide measuring reagent ball.

[0064] Specifically, in this embodiment, the mixed solution includes the following components: Tris buffer solution 5-150mmol / L, magnesium sulfate 0.1-10g / L, bovin...

Embodiment 2

[0066] In this embodiment, the buffer includes glycine-sodium hydroxide buffer, the magnesium salt includes magnesium acetate, the stabilizer includes sucrose, and the excipient includes dextran 10,000 and water-soluble starch. The preparation method of the serum carbon dioxide measurement reagent ball is as follows: a certain amount of glycine-sodium hydroxide buffer solution, magnesium acetate, sucrose, dextran 10,000, water-soluble starch, phosphoenolpyruvate, reduced coenzyme I, apple Acid dehydrogenase, phosphoenolpyruvate carboxylase and water are mixed to form a mixed solution, the droplets of the mixed solution are dropped in liquid nitrogen to form ice balls with a volume of about 2.5ul, and the ice balls are freeze-dried to obtain serum Carbon dioxide reagent balls. And the microfluidic chip is prepared by using the serum carbon dioxide measuring reagent ball.

[0067] Specifically, in this embodiment, the mixed solution includes the following components: glycine-so...

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Abstract

The embodiment of the invention relates to the technical field of medical immune in-vitro diagnosis, and particularly relates to a preparation method of a serum carbon dioxide determination reagent ball and the reagent ball. The method comprises the following steps of: dripping a mixed solution into liquid nitrogen in a liquid drop form, so that liquid drops form an ice ball; and then freezing and drying the ice ball to prepare the serum carbon dioxide determination reagent ball, wherein the mixed solution is prepared from the following components: 5 to 150mmol/L of a buffering solution, 5 to 20g/L of phosphoenolpyruvic acid, 0.1 to 10g/L of reduced coenzyme I, 5 to 20KU/L of malic dehydrogenase, 0.1 to 5KU/L of phosphoenolpyruvate carboxylase, 0.1 to 10g/L of a stabilizer and 10 to 100g/L of an excipient. The excipient with the dosage range can ensure the shape and re-melting solubility of the reagent ball and the activity of reactants is protected; the excipient is good for completely freezing and drying the reagent ball. Namely, the serum carbon dioxide determination reagent ball prepared by the method has relatively good shape and re-melting solubility and can be completely frozen and dried, so that the serum carbon dioxide determination reagent ball has relatively high stability and precision and can realize in-time diagnosis.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of in vitro medical immunodiagnosis, in particular to a method for preparing a serum carbon dioxide measurement reagent ball and the reagent ball. Background technique [0002] 90% to 95% CO in serum or plasma 2 to HCO 3 - It is the main buffer base for the body to maintain acid-base balance, and it is also one of the commonly used indicators for judging acid-base metabolism. blood CO 2 Content determination provides an important basis for judging the progress and curative effect of acid-base balance disorders, and is widely used in clinical practice and is one of the indispensable emergency items. [0003] At present, the enzymatic method used clinically to measure serum carbon dioxide, on the one hand, the reagent solution needs to add a circulatory regeneration system of reducing coenzyme I (NADH), and the stability is poor. On the other hand, its dry powder reagents have po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/527C12Q1/32
CPCC12Q1/527C12Q1/32G01N2333/904
Inventor 牟健汪晨宇周慧欣陈明
Owner GENRUI BIOTECH INC
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