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Stable protein solution, preparation method thereof and detection kit

A protein solution and detection kit technology, applied in biological testing, measuring devices, material inspection products, etc., to achieve the effect of improving stability and preventing denaturation

Inactive Publication Date: 2020-04-17
深圳市蔚景生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods still have certain limitations when applied to occasions that require long-term maintenance of high activity of proteins
For example, commercially available HRP-labeled antibody diluents added with antibody protectants and stabilizers generally only allow HRP-labeled antibodies to be stored stably for half a year at 2-8°C

Method used

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  • Stable protein solution, preparation method thereof and detection kit
  • Stable protein solution, preparation method thereof and detection kit
  • Stable protein solution, preparation method thereof and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] In this example, HRP (Sinopharm Group Chemical Reagent Co., Ltd.)-labeled anti-gastrin-17 antibody (stock solution of HRP-labeled antibody reagent) was used for the test. Carbon monoxide (CO) was passed through the HRP-labeled antibody reagent stock solution at a flow rate of 2-4 bubbles / second for 1 hour to treat the heme in the HRP. Then, the treated stock solution of the HRP-labeled antibody reagent was added to the protein diluent formula 1 shown in Table 1 at a ratio of 1:2000 (volume ratio) to prepare the HRP-labeled antibody dilution solution.

[0051] Table 1: Protein Diluent Recipe 1

[0052] components Dosage per liter Na 2 HPO 4 12H 2 o

5.2g NaH 2 PO 4 2H 2 o

0.62g Sodium chloride 8.5g casein 1g BSA 20g Proclin-300 0.5mL purified water 1000 pH 7.4±0.2

[0053] The prepared HRP-labeled antibody dilution solution was used for accelerated stability test at 37°C, on day 0, day 7 and ...

Embodiment 2

[0073] This example is the same as Example 1, including carbon monoxide (CO) treatment of the stock solution of the HRP-labeled antibody reagent, the difference is that the protein diluent is formulated as the HRP-labeled antibody dilution solution using the protein diluent formula 2 shown in Table 4.

[0074] Table 4: Protein Diluent Recipe 2

[0075]

[0076]

[0077] In the same manner as in Example 1, the prepared HRP-labeled antibody dilution solution was subjected to a 37°C accelerated stability test, and the results are shown in Table 5.

[0078] Table 5: 37°C accelerated stability test results of Example 2

[0079]

[0080]It can be seen from Table 5 that the stock solution of the HRP-labeled antibody reagent in this example was treated with carbon monoxide (CO), and the prepared HRP-labeled antibody dilution solution had no obvious OD value until the 14th day in the accelerated stability test at 37°C. decreased, indicating that carbon monoxide (CO) treatment...

Embodiment 3

[0082] In this example, the stock solution of the HRP-labeled antibody reagent was also treated with carbon monoxide (CO), and the protein diluent formula 2 in Example 2 was used to prepare the HRP-labeled antibody dilution solution, which was a test based on the accelerated stability of Example 2. Accelerated stability is used to quickly screen the best formula, and long-term stability is the final confirmation of the formula, and each performance index can be confirmed more specifically. The long-term stability test is carried out under the environment of 2-8 ℃, and the accuracy (recovery rate), minimum detection limit, precision and linear relationship of the test are investigated.

[0083] 1. Accuracy

[0084] Select 3mL of low-concentration test sample and divide it into 3 parts, each 1mL. Add 0.1mL high-value standard solution (400pg / mL, 800pg / mL) of different concentrations of the analyte to two of the samples, so that the final concentration is between 50-1000pg / mL, a...

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Abstract

The invention provides a stable protein solution. The protein solution comprises a buffer solution and a protein containing a heme auxiliary group or a protein conjugate of the protein containing theheme auxiliary group and other proteins. Ferrous ions in the heme auxiliary group are combined with carbon monoxide, so that the protein or the protein conjugate can be prevented from being oxidized and denatured in the storage process, and the stability of the protein solution is improved. The protein solution may also include other stabilizers. The protein solution disclosed by the invention canbe stably stored for at least one year at 2-8 DEG C, has good stability, and can be used for preparing a chemiluminescence detection kit or an enzyme-linked immunosorbent assay kit, such as a detection kit for detecting gastrin 17, pepsinogen I and pepsinogen II.

Description

technical field [0001] The invention relates to the technical field of biochemical detection, in particular to a stable protein solution, wherein the protein is a protein containing a heme prosthetic group or a protein conjugate of a protein containing a heme prosthetic group and other proteins, and the invention also relates to the protein A method for preparing the solution, and a detection kit containing the protein solution, such as a chemiluminescent detection kit or an enzyme-linked immunological detection kit. Background technique [0002] Heme is an important component of some proteins, such as hemoglobin, myoglobin, cytochrome C oxidase, catalase and peroxidase, which all contain heme. These proteins have important physiological functions in organisms. Heme is a metalloporphyrin complex formed by ferrous (II) ion and protoporphyrin IX (see the heme structural formula shown below). The ferrous iron (II) ion is bound by the four nitrogen atoms of the porphyrin ring w...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/573G01N33/53
CPCG01N33/74G01N33/573G01N33/5306G01N2333/595G01N2333/96477
Inventor 董婷王亚磊
Owner 深圳市蔚景生物科技有限公司
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