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A chemiluminescence reagent and reagent technology, which is applied in the direction of chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problems of affecting HRP activity, affecting detection sensitivity, linearity, skimmed milk powder and casein perishable, etc.
Active Publication Date: 2016-08-10
HELIOSENSE BIOTECH INC +1
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Problems solved by technology
Some documents (Anal.Biochem.1995,231,170-174 and EP0650044A2) have reported the use of skim milk powder, casein, monosaccharides, ionic detergents, etc., but the storage stability of skim milk powder and casein, batch The difference between batches is a huge problem. In the absence of preservatives, skim milk powder and casein are easy to deteriorate; different batches of skim milk powder and casein may contain different amounts of protein, so poor quality between batches is not easy control
In addition, traditional chemiluminescence techniques generally use chelating agents (such as EDTA, etc.) to reduce the background, but they can only meet part of the requirements. Too high EDTA will seriously affect the activity of HRP, thereby affecting the detection sensitivity, linearity, etc.
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Embodiment example 1
[0063] i) Implementation case 1, formula containing antioxidant acetylcysteine:
[0064] a) Preparation of liquid A
[0065] Luminol monosodium salt, the concentration is 0.01%;
[0066] Phenothiazin-10-yl-propylsulfonic acid sodium salt, the concentration is 0.05%;
[0067] Acetylcysteine at a concentration of 0.00003%;
[0068] Prepared in pure water.
[0069] b) Preparation of liquid B
[0070] Sodium perborate, the concentration is 0.03%;
[0071] Disodium edetate, the concentration is 0.008%;
[0072] It is formulated in a sodium tetraborate-boric acid buffer solution with a pH value of 8.5.
Embodiment example 2
[0073] ii) Implementation case 2, formula containing antioxidant vitamin B1:
[0074] a) Preparation of liquid A
[0075] Luminol monosodium salt, the concentration is 0.01%;
[0076] Phenothiazin-10-yl-propylsulfonic acid sodium salt, the concentration is 0.05%;
[0077] Vitamin B1 at a concentration of 0.001%.
[0078] Prepared in pure water.
[0079] c) Preparation of liquid B
[0080] Sodium perborate, the concentration is 0.03%;
[0081] Disodium edetate, the concentration is 0.008%;
[0082] It is formulated in a sodium tetraborate-boric acid buffer solution with a pH value of 8.5.
[0083] B) Application of HRP chemiluminescence substrate in enhanced chemiluminescence enzyme immunoassay
[0084] The following is an example of an enhanced chemiluminescence enzyme immunoassay for hepatitis B antigen (HBeAg or HBcAg) in a 96-well plate to describe the operation plan:
[0085] 1. Detection of hepatitis B e antigen HBeAg (hepatitis B e antigens)
[0086] 1. Coat a 9...
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Abstract
The invention discloses a chemiluminescence reagent, and relates to a chemiluminescence reagent which is mainly applied to a biotechnology, bioscience, medical diagnosis, food safety, and environment-friendly detection. The chemiluminescence reagent is prepared from the materials by mass percent: at least one 0.001-0.1% luminous reagent, at least one 0.001-0.2% oxidant, at least one 0.01-0.5% luminous enhancer, at least one 0.000001-0.05% background control agent, and a buffer solution of which the pH value is 8-11. The key is as follows: chemiluminescence is adjusted by utilizing the background control agent (particularly the antioxidant); the chemiluminescence reagent is valid on background control of enhanced chemiluminescence enzyme immunoassay detection, improvement of the signal to noise ratio and the like. The basis is how to control luminol free radical and concentration of related free radical in a horse radish peroxidase-luminol luminescence system.
Description
technical field [0001] The invention relates to a chemiluminescence reagent, in particular to a chemiluminescence reagent mainly used in biotechnology, life science, medical diagnosis, food safety and environmental protection detection. Background technique [0002] Chemiluminescence refers to a phenomenon in which visible light is generated during a chemical reaction. It usually refers to the fact that some compounds absorb chemical energy and are excited from the ground state to the excited state by absorbing chemical energy, and then release energy when de-exciting. Photons are generated to realize the transformation from chemical energy to light energy, resulting in the phenomenon of luminescence. By measuring the light intensity of the chemiluminescence reaction, the content of trace compounds can be analyzed. The chemiluminescent immunoassay (CLIA) developed in recent years is the product of the combination of chemiluminescent technology and immunological methods. It ...
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