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The difference between acridinium ester direct chemistry and alkaline phosphatase (AP) chemiluminescence
Chemiluminescence immunoassay methods can be roughly divided into three categories according to their labels, direct chemiluminescence, enzymatic chemiluminescence, and electrochemiluminescence. Alkaline phosphatase (AP) chemiluminescence is a kind of enzymatic chemiluminescence, which is specially developed for the application of alkaline phosphatase (AP) enzyme-linked immunosorbent assay. It is recommended for enzyme-linked immunosorbent assay and western blotting. Method, Southern blotting or any other solid surface for staining application and detection of alkaline phosphatase-labeled nucleotide and protein probes.
The biggest difference between direct chemiluminescence of acridinium ester and alkaline phosphatase (AP) chemiluminescence is that acridinium ester is direct chemiluminescence, which is hydrolyzed under the action of hydrogen peroxide and hydroxide to emit light, which belongs to the flash type (luminescence value is about 0.5s Enter the highest value, the light will end within 1s). Alkaline phosphatase catalyzes the luminescence of substrates such as AMPPD, and belongs to the glow type (entering the luminescence plateau period, continuous luminescence). Therefore, the direct chemiluminescence of acridinium esters has higher requirements for the instrument. It is necessary to add the excitation solution in situ and complete the reading within 1s.
Desheng acridine ester chemiluminescence reagent
One disadvantage of alkaline phosphatase is that the enzyme and the substrate need to be incubated and reacted for a period of time before the luminescence value can reach the plateau. In particular, AMPPD needs to incubate for 5 minutes to reach the luminescence plateau. Compared with the direct chemiluminescence of acridinium esters in 1 second, it will affect the detection speed. Another new type of alkaline phosphatase substrate, Lumigen APS-5 can reach the luminescence plateau within 30 seconds, but because of its sensitivity to temperature and poor long-term stability than AMPPD, it has not been widely used in chemiluminescence systems. . As a leader in alkaline phosphatase chemiluminescence, Beckman has solved the stability problem of APS-5 and applied for a global patent this year. Perhaps in the near future, Beckman will switch the substrate to a more advanced APS-5 to enhance competitiveness.
To establish a new chemiluminescence immunoassay platform, it is necessary to choose a suitable type of luminescence, whether it is direct chemiluminescence, enzymatic chemiluminescence, or electrochemiluminescence. From the current point of view, most companies choose alkaline phosphatase, or acridinium ester direct chemiluminescence. From the perspective of reagent development, acridinium esters may have a slight advantage over alkaline phosphatase, but on the whole, there is no obvious gap between the two. Giants such as Beckman and Mindray use alkaline phosphatase. , Can also gain a leading position in the mainstream market.
Therefore, the choice of chemiluminescence platform is a question that needs to be considered comprehensively. It needs to consider the availability of talents, the comprehensive difficulty of reagents and instruments, and the input cost, reagent cost, etc. Currently, Desheng mainly uses acridinium esters. Direct chemiluminescence method, its product advantages are shown in the aspects of good hydrolytic stability, high purity, good thermal stability and high sensitivity, and after use by customers, it is unanimously regarded as an ideal chemical marker.