Method for detecting target by using chemiluminescence enhancer modified antibody
A chemiluminescence and luminescence enhancement technology, applied in the field of immunodetection, can solve the problems of special luminescence substrate, unfavorable for promotion and application, difficult synthesis, etc. sufficient effect
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Embodiment 1
[0040] The preparation of embodiment 1 phenoxazine activated ester
[0041] 0.27 g (1 mMol) of phenoxazin-10-yl-acetic acid, 0.17 g (1.5 mMol) of N-hydroxysuccinimide (NHS), and 0.31 g (1.5 mMol) of dicyclohexylcarboimide (DCC) were mixed. Add 2 ml of anhydrous DMSO. After shaking to dissolve, react at room temperature for 36 hours. The activated ester of the phenoxazine is obtained. The activated ester can be used for a long time after freezing.
Embodiment 2A
[0042] Example 2 AFP primary antibody labeled phenoxazine
[0043] Add 1.3 microliters of the activated ester prepared in Example 1 to the solution of AFP primary antibody (containing 10 mg of mouse monoclonal antibody). The molar ratio of activated ester to antibody was 10:1. The two were reacted at room temperature for 6 hours and then dialyzed three times (in physiological saline). The obtained phenoxazine-labeled antibody solution was diluted with physiological saline to a concentration of 100 ug / ml for use.
Embodiment 3
[0044] The composition of embodiment 3 test kits
[0045] R1 reagent: containing 2ug / ml AFP primary antibody (phenoxazine-labeled), containing 2ug / ml AFP secondary antibody (HRP enzyme-labeled), containing 0.1% concentration of luminol, the medium is 0.1M PBS pH=7.4.
[0046] R2 Reagent: Contains 0.1% hydrogen peroxide.
[0047] Chemiluminescence detection instrument: LUMO chemiluminescence immunoassay analyzer produced by Antu Company
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