Alkaline phosphatase-catalyzed chemiluminescent substrate enhancer and application thereof

Abstract

The invention discloses an alkaline phosphatase-catalyzed chemiluminiscence substrate enhancer and an application thereof. The alkaline phosphatase-catalyzed chemiluminiscence substrate enhancer comprises a fluorescent compound and a surfactant, the fluorescent compound is water-soluble quantum dots, the surfactant is a Gemini surfactant, and a chemiluminiscence substrate solution containing the enhancer comprises 50-500 mM of 2-amino- 2-methyl-1-propanol, 0.1-1 mM of AMPPD, 1-10 mM of MgCl2, 10-30 mg / L of the enhancer and 100-1000 mg / L of preservative. The Gemini surfactant is creatively selected to be matched with the water-soluble quantum dots to obtain the alkaline phosphatase-catalyzed chemiluminescence substrate enhancer, a distance between the water-soluble quantum dots and the AMPPD substrate is shortened by utilizing an internal hydrophobic microenvironment of micelles formed by the Gemini surfactant, and then light energy transfer is generated; and a quantum yield of the water-soluble quantum dots is far higher than that of chemiluminescence molecules so that a luminescence value of the chemiluminescence substrate liquid containing the enhancer is remarkably enhanced, andperformances of luminescence value measurement, such as precision, sensitivity and luminescence rate stability, are further increased.

Description

technical field [0001] The invention belongs to the technical field of immunoassay, and in particular relates to an alkaline phosphatase-catalyzed chemiluminescence substrate enhancer and application thereof. Background technique [0002] Chemiluminescence is a kind of light radiation phenomenon accompanied by the chemical reaction of substances, which can be divided into direct luminescence and indirect luminescence. Direct luminescence is the simplest chemiluminescence reaction, which consists of two key steps: excitation and radiation. For example, two substances A and B undergo a chemical reaction to form C substance, the energy released by the reaction is absorbed by the molecules of C substance and transitions to the excited state C*, and the excited C* generates light radiation in the process of returning to the ground state. Here C* is a luminescent body, and because C directly participates in the reaction in this process, it is called direct chemiluminescence. Ind...

AI Technical Summary

Problems solved by technology

[0003] 1,2-dioxetane compound (AMPPD) is an ultrasensitive alkaline phosphatase substrate, under the catalysis of alkaline phosphatase (AP), the phosphate group is hydrolyzed to form an unstable Intermediate, self-decomposing, -O-O- four-membered ring breaks and releases a large amount of energy, thereby stimulating chemiluminescence reaction and emitting photons. Although its signal can last for more than 20 hours, its luminescence rate is low. Commercially available chemiluminescence substrates The solution is a substrate solution compounded with an enhancer and AMPPD to increase the luminous intensity

It is found in the prior art that enhancers that can be used for luminescent substrates mainly include globular proteins such as bovine serum albumin, and cationic surfactant enhancers such as polyethylene polymers with quaternary phosphonium side groups, but they are expensive and require However, adding too much surfactant will easily produce too much foam, which will affect the accuracy of luminescence value detection; meanwhile, the enhancement factor of the above-mentioned enhancer is low, which will lead to low precision of luminescence value measurement and low luminescence rate. Uniformity and other issues

Images