An Enzymatic Chemiluminescent Substrate for Alkaline Phosphatase

An enzymatic chemiluminescence and phosphatase technology, which is applied in the field of immune technology detection, can solve the problems of low luminous intensity and slow light up, and achieve the effects of high sensitivity, long duration, thermal stability and real-time stability

Active Publication Date: 2018-04-20
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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  • An Enzymatic Chemiluminescent Substrate for Alkaline Phosphatase
  • An Enzymatic Chemiluminescent Substrate for Alkaline Phosphatase
  • An Enzymatic Chemiluminescent Substrate for 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 3, compare enzymatic chemiluminescent substrate (APSH) of the present invention and Beckman chemiluminescent substrate AccessSubstrate to determine 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 of TSH...

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Abstract

The present invention relates to an alkaline phosphatase enzymatic chemiluminescent substrate. The enzymatic chemiluminescent substrate uses water as a solvent, and also comprises the following components: 2-amino-2-methyl-1-propanol; AMPPD; a luminescence enhancer which is sodium fatty alcohol polyoxyethylene ether carboxylate coupled with a fluorescent compound. The luminescence enhancer has a co-surfactant effect, allowing the complex to be better combined into a chemiluminescent buffer system, thereby significantly increasing the chemiluminescent efficiency. The alkaline phosphatase enzymatic chemiluminescent substrate has advantages such as high strength, high sensitivity, a long duration and good stability, and fully satisfies clinical detection requirements. A main performance indicator of the chemiluminescent substrate reaches a foreign product level, and the costs of the chemiluminescent substrate are significantly reduced.

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 Patents(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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