Strongly Chemiluminescence reagent Acridinium Esters Invitro Immunoassays Using

Acridine ester is a chemical substance that can be used as a chemiluminescent label. Acridine salts and related compounds have been widely proven to be very useful chemiluminescent labels. Its stability, labeling specificity and detection sensitivity surpass that of radioisotopes. . Under alkaline conditions, NHS is substituted as a leaving group, and the protein forms a stable amide bond with the acridine ester. However, acridinium ester is only a general term for acridinium salt. If subdivided, Desheng can provide 7 different acridinium esters (see the table below) for you to choose from, so what are these 7 acridinium esters? What are the similarities and differences? Let us continue to look down.

1.1 Acridinium Ester No. 0-Acridinium Ester (AE-NHS)

【CAS】None

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】N/A

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

acridinium ester powder

1.2 Acridinium Ester No. 1-Acridinium Ester (DMAE-NHS)

【English name】2',6'-dimethyl-4'-(N-succinimidyloxycarbonyl)phenyl-10-methyl-acridinium-9-carboxylate methosulfate

[Chinese name] 9-[[4-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]-2,6-dimethylphenoxy]carbonyl]-10-methyl Acridine Methyl Sulfate

【CAS】115853-74-2

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】594.13, C29H26N2O10S

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

1.3 Acridinium Ester No. 2-Acridinium Ester (Me-DMAE-NHS)

【English name】2',6'-Dimethylcarbonylphenyl 10-Methyl-9-acridinecarboxylate 4'-NHS Ester Triflate

[Chinese name] 2',6'-dimethylcarbonylphenyl 10-methyl-9-acridine carboxylate 4'-NHS ester trifluoromethanesulfonate

【CAS】None (115853-74-2, DMAE-NHS analogue)

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】632.56, C29H23F3N2O9S

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

1.4 Acridinium Ester No. 3-Acridinium Ester (NSP-DMAE-NHS)

【English name】2',6'-DiMethylcarbonylphenyl-10-sulfopropylacridinium-9-carboxylate 4'- NHS Ester

[Chinese name] 9-[[4-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]-2,6-dimethylphenoxy]carbonyl]-10-( 3-

Sulfopropyl)-acridine inner salt

【CAS】194357-64-7

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】590.6, C30H26N2O9S

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

1.5 Acridine ester No. 4-acridine salt (NSP-SA)

[English name] 3-[9-(((3-(carboxypropyl)[4-methxylphenyl]sulfonyl)amine)carboxyl]-10-acridiniumyl)-1-propanesulfonateinner salt

[Chinese name] Acridine salt (NSP-SA)

【CAS】211106-69-0

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】584.66, C28H28N2O8S2

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

1.6 Acridine ester No. 5-acridine salt (NSP-SA-NHS)

[English name] 3-[9-(((3-(N-succinimidyloxycarboxypropyl)[4-methxylphenyl]sulfonyl)amine)carboxyl]-10-acridiniumyl)-1-propanesulfonateinner salt

[Chinese name] Acridine salt (NSP-SA-NHS)

【CAS】199293-83-9

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular Weight】681.73, C32H31N3O10S2

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

1.7 Acridine ester No. 6-acridinium hydrazide (NSP-SA-ADH)

【CAS】None

【Appearance】Yellow solid or powder

【Purity】≥98%

【Molecular weight】740.85, C34H40N6O9S2

【Storage conditions】-20℃, avoid light

[Transportation Conditions] It can be transported at room temperature (20-25°C) within 48 hours, and it needs to be transported at -20°C for more than 48 hours.

two. The structural difference of acridine esters

Six kinds of acridine esters, of which Nos. 1 to 3 are acridine esters; Nos. 4 to 6 are acridine sulfonamide lactates; Nos. 1 to 4 are NHS active esters; No. 5 is acridine carboxylic acid with carboxyl group; 6 The number is acridine hydrazide, which contains free amino groups.

three. Hydrolysis resistance and stability

Traditional acridinium ester No. 0 (AE-NHS) and No. 1 to 3 have been modified in their structure to increase steric hindrance and enhance hydrolysis resistance. No. 0 is only stable in acidic solutions. When the pH value is greater than 6.3, it is easy to hydrolyze, while No. 1 to 3 are not. For example, it is stable in PB buffer with pH 7.0 at room temperature. After 16 days, the luminescence activity only decreases. 3.6%.

Nos. 4 to 6 have introduced sulfonamide groups, which are more stable than acridine esters (No. 0 to 3). The reason is that the C-N bond and the C-O bond have different bond levels, and the C-N bond is larger than the C-O bond. Acridine amide compounds (No. 4 to 6) are also stronger against hydrolysis than acridine ester compounds (No. 0 to 3). No. 4～6 are stable in acidic solution (pH<4.8). When stored with protein conjugates at room temperature for 4 weeks, the photon yield will not decrease. Freeze-dried products can be stored for more than one year at -20°C. .

four. Hydrophilicity

Nos. 3 to 6 are modified in hydrophilicity on the basis of Nos. 1 to 2. The introduction of propanesulfonic acid internal salt increases its water solubility and makes the markers formed with antibodies, nucleic acids, etc. water soluble , The subsequent luminescence detection can be performed in water.

Fives. The difference in marking

Since the essence of the antibody is a protein, it contains an amine group and can directly react with No. 1 to No. 4 (NHS active ester) for coupling.

No. 5 is acridine carboxylic acid, which requires the addition of condensing agent EDCI, etc., to have a coupling reaction with amine-containing proteins.

No. 6 is acridine hydrazide, which contains free amino groups. The end of acridine hydrazide is suitable for direct coupling to polysaccharides, nucleic acids or proteins containing aldehyde groups.

six. Comparison of luminous performance

As acridines No. 1 to No. 6 have the same luminescence matrix and mechanism, their luminescence performance should not be much different.