Advantages of acridine esters compared to other chemiluminescent substrates
Release time:
2021-04-12
Acridinium ester chemiluminescence reagent is an advantageous reagent in tubular chemiluminescence. Compared with luminol, AMPPD, ruthenium terpyridine and other chemiluminescence substrates, it has unique advantages. It has ultra-high sensitivity and operation Simple and easy to automate operation.
The chemiluminescence reaction of acridinium ester does not require a catalyst:
Different acridinium ester chemiluminescent substrates
When acridine esters (including acridine sulfonamide) are oxidized by hydrogen peroxide under alkaline conditions, they emit light with a wavelength of 470nm and have high luminous efficiency. The excited state product N-methylacridone is the luminescence The luminous body of the reaction system. It does not require enzyme catalysis and directly participates in the luminescence reaction.
Acridine esters can be directly combined with antibodies or magnetic beads:
In the immunoassay, the antibody (antigen or magnetic beads) is directly labeled with acridinium ester, and after the immunoreaction with the corresponding antigen (antibody) in the sample to be tested, a solid-phase coating antibody-the antigen to be tested-acridinium ester label is formed The antibody complex only needs to add oxidant (hydrogen peroxide) and sodium hydroxide to make it into an alkaline environment after washing, and the acridinium ester decomposes and emits light without a catalyst.
Luminol and AMPPD need enzymes to participate in the luminescence:
This kind of luminescent substrate belongs to enzymatic reaction. Luminol needs peroxidase POD or HRP to catalyze luminescence, while AMPPD needs alkaline phosphatase to catalyze luminescence reaction. Since enzymes are added to the reaction, and the enzyme activity is affected by temperature and environmental pH, the reaction system is relatively complicated. In addition to the pH required for the labeled protein, the appropriate pH of the enzyme needs to be considered.
The cost of using ruthenium terpyridine is high:
This luminescent substrate belongs to electrochemiluminescence, and has the advantages of rapid detection, controllable reaction conditions, wide detection range, high precision and sensitivity, and low sample detection dosage. But the disadvantages are: the flow colorimetric cell used in the instrument, cross-contamination is a potential problem; it is too sensitive to environmental factors and other non-specific reactions; the detection instrument is expensive and is less used in China.
In summary, the advantages of acridine ester luminescent agents are that they do not require enzyme catalysis and no enhancer, thereby reducing background luminescence, improving signal-to-noise ratio, less interference, and the use and development costs are much lower than electrochemiluminescence. There are six kinds of acridine esters developed by Desheng, including DMAE-NHS, Me-DMAE-NHS, NSP-DMAE-NHS, NSP-SA-NHS, etc.
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