Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase

An enzymatic chemiluminescence and phosphatase technology, applied in the field of immunological detection, can solve the problems of slow light emission and low luminescence intensity, and achieve the effects of long duration, high sensitivity and cost reduction.

Active Publication Date: 2015-10-21
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the chemical reaction of AP catalyzing AMPPD is a hydrolysis reaction. Although it has a long plateau period, its luminous intensity is low and the light emission is slow. Therefore, this chemical reaction still needs to be optimized.

Method used

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  • Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase
  • Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase
  • Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the preparation of enzymatic chemiluminescent substrate (APSH)

[0038] (1) Materials: 2-amino-2-methyl-1-propanol, AMPPD, MgS0 4 and ZnCl 2 All are commercially available and chemically pure; ProClin300 is a product of Supelco Company in the United States; the luminescence enhancer HOBEP-1 is obtained by coupling fatty alcohol polyoxyethylene ether carboxylate sodium with coumarin fluorescent compounds, and the luminescence enhancer HOBEP-2 is It is obtained by coupling sodium fatty alcohol polyoxyethylene ether carboxylate with fluorescein-based fluorescent compounds, and the luminescence enhancer HOBEP-3 is obtained by coupling sodium fatty alcohol polyoxyethylene ether carboxylate with naphthalimide fluorescent compounds. The enhancer HOBEP-4 is obtained by coupling sodium fatty alcohol polyoxyethylene ether carboxylate with rhodamine fluorescent compounds, and the luminescence enhancer HOBEP-5 is obtained by coupling sodium fatty alcohol polyoxyethyl...

Embodiment 2

[0073] Example 2. Comparison of the enzymatic chemiluminescence substrate (APSH) and the Beckman chemiluminescence substrate (Access Substrate) in this example to test the AP signal intensity and linear relationship at each gradient concentration

[0074] The chemiluminescent substrate ASPH-1 prepared according to Table 1.2, the chemiluminescent substrate ASPH-2 prepared according to Table 2.2, the chemiluminescent substrate ASPH-3 prepared according to Table 3.2, and the chemiluminescent substrate prepared according to Table 4.2 were used respectively. The chemiluminescent substrate ASPH-4, the chemiluminescent substrate ASPH-5 prepared according to Table 5.2, and the commercially available chemiluminescent substrate Access Substrate of Beckman Company of the United States tested the signal intensity under the conditions of each gradient concentration AP (relative luminescence intensity RLU) and linear relationship, the test results are shown in Table 6 below.

[0075] Table ...

Embodiment 3

[0086] Embodiment three, compare enzymatic chemiluminescent substrate (APSH) of the present invention and Beckmann chemiluminescent substrate Access Substrate to measure the minimum detection limit (LOD) of thyroid-stimulating hormone (TSH) quantitative detection kit (magnetic particle chemiluminescence method) )

[0087] The chemiluminescent substrate ASPH-1 prepared according to Table 1.2, the chemiluminescent substrate ASPH-2 prepared according to Table 2.2, the chemiluminescent substrate ASPH-3 prepared according to Table 3.2, and the chemiluminescent substrate prepared according to Table 4.2 were used respectively. The minimum detection of chemiluminescent substrate ASPH-4, chemiluminescent substrate ASPH-5 prepared according to Table 5.2 and the commercially available chemiluminescent substrate Access Substrate from Beckman Company of the United States for the determination of thyroid-stimulating hormone (TSH) quantitative detection kit limit (LOD).

[0088] The content...

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Abstract

The invention relates to an enzyme-promoting chemiluminiscence substrate using alkaline phosphatase. Using water as solvent, the enzyme-promoting chemiluminiscence substrate comprises 2-amino-2-methyl-1-propyl alcohol, AMPPD, and a luminescence enhancer link-coupled by sodium fatty alcohol polyoxyethylene ether carboxylate and fluorescent compounds. According to the enzyme-promoting chemiluminiscence substrate using alkaline phosphatase, the luminescence enhancer can achieve the effect of co-surfactant, the compounds can be better combined into a chemiluminiscence buffer system, and thus the chemiluminiscence efficiency is greatly improved. The enzyme-promoting chemiluminiscence substrate using alkaline phosphatase has the advantages of being high in strength, high in sensitivity, long in duration, good in stability and the like. Clinical testing requirements can be met completely, the main performance index has already reached the level of foreign products, and the cost of the chemiluminiscence substrate is greatly lowered.

Description

technical field [0001] The invention relates to the field of immunotechnical detection, in particular to a high-efficiency enzymatic chemiluminescent substrate that can be used for chemiluminescent immunodetection. Background technique [0002] Chemiluminescence refers to the generation of an excited state product (C * ), in the process of returning to the ground state, the released energy is converted into photons (energy hγ), thereby producing luminescence. [0003] Chemiluminescent immunoassay is an ultra-high-sensitivity micro-detection technology developed after enzyme immunoassay, radioimmunoassay, and fluorescence immunoassay. It has high sensitivity, wide detection range, simple and fast operation, good stability of markers, and no pollution. And other advantages, so it has become a non-radioactive immunoassay technology that develops very rapidly in the world, and is the most ideal method for immunoquantitative analysis at present. [0004] In chemiluminescent imm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/535
CPCG01N21/76G01N33/53
Inventor 秦枫马竹凤王静张伟李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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