Among various non-radioimmunoassay methods, chemiluminescence immunoassay (CLIA) has the advantages of high sensitivity, wide detection range, good precision, stable markers, full automation, and a wide range of applications. It is based on acridine ester derivatives. The characteristics of CLIA, which is a luminescent label, are: simple labeling, stable labeling, no need for catalysts, low natural background, few external interference factors, high signal-to-noise ratio, and simple luminescent system.
Acridine ester is a general term for acridine salt, which refers to a class of chemical substances that can be used as chemiluminescent markers. There are many different series of acridine esters, including acridine salt DMAE-NHS, acridine ester NSP-DMAE- NHS, acridine salt NSP-SA, etc. Next, we will introduce the synthesis method, performance and advantages of acridine ester DMAE-NHS.
Synthetic route of acridine ester DMAE-NHS:
1. Dissolve 4-hydroxy-3,5-dimethylbenzoic acid and N-hydroxysuccinimide in N,N-dimethylformamide (DMF), and react under the action of a condensing agent to obtain a crude product , The pure product is obtained after recrystallization;
2. Dissolve the product 1-((4-hydroxy-3,5-dimethylbenzoyl)oxy)-2,5-pyrrolidinedione and acridine-9-carbonyl chloride in anhydrous pyridine In the process, the crude product is obtained by heating the reaction under the action of the catalyst, which is separated and purified by column chromatography.
3. Dissolve the product obtained in 2 and dimethyl sulfate in anhydrous toluene to react, filter the obtained mixture, wash the filter residue and then recrystallize to obtain the target product of acridinium ester chemiluminescent substrate DMAE-NHS.
Introduction of DMAE·NHS's luminous performance
1. Research on the luminescence performance of DMAE-NHS shows that the luminescence of DMAE-NHS is flash, the luminous intensity reaches the maximum at 0.4s, the luminous half-life is about 0.9s, and the luminous intensity is 6.11×1018cps/mol. The composition of the luminescence initiating reagent has an influence on its luminescence intensity, and surfactants such as CTAC, TritonX-100, Tween-20, etc. can enhance its luminescence.
2. The luminescence properties of DMAE-NHS and antibody conjugate DMAE-NHS-Ab are similar. When the molecular ratio of DMAE-NHS to Ab is 2.4, the luminescence intensity is 1.8×1019cps/molAb. DMAE-NHS and DMAE-NHS-Ab have good thermal stability. At 4°C or frozen, in the PB buffer at pH 7.0, their luminescence activity or immunological activity basically does not decrease after half a year, room temperature or 37°C Under the condition that the luminescence activity is reduced due to hydrolysis, their thermal stability decreases with the increase of temperature, and decreases with the increase of the pH value of the melt.
Introduction to the advantages of DMAE-NHS
The research results show that DMAE-NHS has good luminescence properties, is easy to couple with antibodies, the conjugate is stable, and the luminescence system is simple. It is an ideal chemiluminescence immunoassay marker, which is established by using DMAE-NHS as a luminescence marker. TSHCLIA, TSHBAS-CLIA and TT4BAS-CLIA methods, standard curve linearity, high signal-to-noise ratio, wide detection range, high sensitivity, good precision, accuracy and soundness, and IRMA method and CibaComingCLIA clinical measurement The value correlation is good.
The acridinium ester luminescence system has obvious advantages and has better sensitivity in in vitro diagnosis. At present, the application of acridinium ester chemiluminescence immunoassay technology has gradually developed into a trend in the fields of medical testing, diagnostic analysis, environmental testing, and food testing.  Desheng Technology provides reliable acridine ester and acridine salt products, which can meet the scientific research and production needs of scientific research institutions and kit manufacturers.