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Introduction of chemiluminescent agents commonly used in chemiluminescence immunoassays
2019-06-26
来源：亚科官网
Chemiluminescence immunoassay (CLIA) is a type of immunoassay that directly labels antigens or antibodies with chemiluminescent agents. Acridinium ester is an ideal luminescent substrate which can be oxidized by hydrogen peroxide in an alkaline environment. It is the fastest immunoassay method currently developed and popularized, and it is also the most advanced labeling immunoassay technology. The sensitivity and accuracy are several orders of magnitude higher than that of enzyme-free and fluorescent methods. It can completely replace radioimmunoassay and completely eliminate ELISA. It has the advantages of high sensitivity, strong specificity, low reagent cost, stable reagent and effective period (6-18 months), stable and rapid method, wide detection range, simple operation and high automation.
The chemiluminescent agent used as a marker should meet the following conditions:
1. participate in the chemiluminescence reaction.
2. a stable conjugate reagent can be formed after coupling with an antigen or antibody.
3. high quantum effects and reaction kinetics remain after coupling.
4. should not be altered or minimally altered the physical and chemical properties of the label, especially the immunological activity,.
Luminol and acridine ester luminescent agents are commonly used as marker luminescent agents.
Classification of markers used in chemiluminescence immunoassays
1 Direct chemiluminescent agent: The antigen or antibody is directly labeled with a chemiluminescent agent.
Acridine esters are currently commonly used direct labeling luminescent agents.
(1). Acridine ester: When oxidized by H2O2 under alkaline conditions, light having a wavelength of 470 nm is emitted, which has a high luminous efficiency, and the excited state product N-methylacridone is an illuminant of the luminescent reaction system. Acridine ester luminescent agents can be directly used to label haptens, proteins, etc. When used for labeling antibodies, the binding is stable, and the binding does not reduce the generation of photons, and high specific activity can be obtained, which is beneficial to the establishment of two-site chemiluminescence immunoassay.
2. Terpyridine pyridinium: terpyridine pyridinium is an electrochemical luminescent agent, and the electron donor tripropylamine (TPA) on the surface of the anode electrode can simultaneously lose an electron and oxidation reaction.
2. Enzymatic reaction illuminator. The luminescent agent (substrate) is caused to emit light by the catalytic action of a labeling enzyme (horseradish peroxidase and alkaline phosphatase).
The horseradish peroxidase-catalyzed luminescent agent is luminol and its derivatives; the alkaline phosphatase-catalyzed luminescent substrate is AMPPD.
(1) Luminol and its derivatives. Luminol, isoluminol and its derivatives have chemiluminescent properties. Luminol is the earliest synthetic luminescent substance.
(2) AMPPD is a new chemiluminescent agent with two important parts in its molecular structure. One is a dioxet ring connecting a benzene ring and adamantane, which can break and emit photons. The other is a phosphate group. It maintains the stability of the entire molecular structure.
Related links: acridinium ester
Edited by Suzhou Yacoo Science Co., Ltd.
