The chemiluminescent agent used for luminescence is very different!
October 23 , 2024
Chemiluminescence immunoassay technology has long been well-known in the field of in vitro diagnostics! These include, direct chemiluminescence, electrochemiluminescence, enzymatic chemiluminescence, indirect chemiluminescence. So many types of luminescence use luminescent agents that are also very different! 1. Acridine ester
Direct chemiluminescence
Acridine ester (AE) is a tricyclic organic compound that is susceptible to oxidation. When oxidized by H2O2 under alkaline conditions, it emits light with a wavelength of 470nm and has a high luminous efficiency. Its excited product N-methylacridone is the luminite of the luminescence reaction system. It is used for the labeling of haptens and proteins or polyclonal or monoclonal antibodies.
It is a synthetic substance, which does not exist in nature, so there is no background noise interference; Acridine ester is relatively insensitive to changes in pH and temperature. At the same time, its molecular weight is only 481, the spatial hindrance effect is small, and the oxidation reaction does not need a catalyst, as long as it can be carried out in an alkaline environment.
The luminescence reacts quickly and has a low background. During the oxidation reaction, the conjugate is decomposed, so the luminescence of free acridine ester is not inhibited, and the reagent has good stability. It is an excellent choice of luminescent agent in the industry.
Terapridine ruthenium [RU(bpy)3]2+ is an electrochemiluminescent agent that oxidizes with the electron donor tripropylamine (TPA) on the surface of the anode. Direct labeling of antigens or antibodies.
The luminescence is characterized by a stable light emission of only 0.01 ms, reaching a peak at 300 ms, and the electrochemiluminescence cycle of hundreds of thousands of cycles per second greatly improves the sensitivity of the analysis.
The marking is stable and easy to control, but the electric field requires a special measuring cell, and it takes 40-60 seconds to clean the measuring cell after each measurement to restore the electrode to the state, which is easy to cause cross-contamination, affect the accuracy of the results, and reduce the analysis speed.
3.Luminol and its derivatives
Enzymatic chemiluminescence
Luminol (3-aminobenzoylhydrazide), isoluminol (4-aminobenzoylhydrazide) and their derivatives have chemiluminescent properties.
In neutral solution: usually present as dipole ions (zwitterions).
In alkaline solution: it becomes a divalent anion, which can be oxidized by oxygen molecules into an intermediate that can produce chemiluminescence phenomenon.
In oxidants: luminol is converted to an excited state, which decays to the ground state and fluoresces.
4. adamantane AMPPD
Enzymatic chemiluminescence
In general, adamantane structure is very stable, but when alkaline phosphatase is encountered, alkaline phosphatase will catalyze adamantane to remove the phosphate group to form an unstable intermediate, which will automatically decompose and produce a 470nm light signal, which can last for tens of minutes to several hours.
5. Photoinduced chemiluminescence
peculiarity
Short reaction time: A homogeneous reaction pattern.
High sensitivity: The reaction process is amplified step by step, and the luminescence rapidly improves the sensitivity.
Good stability: The whole energy (light) generation, transfer and amplification process is very stable, and it is not easily affected by pH, ionic strength and temperature.